Adapta&on and Design of Panels for the CyTOF. December 5, 2014 CyTOF User Mee&ng University of Virginia Mike Leipold

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Adapta&on and Design of Panels for the CyTOF. December 5, 2014 CyTOF User Mee&ng University of Virginia Mike Leipold"

Transcription

1 Adapta&on and Design of Panels for the CyTOF December 5, 214 CyTOF User Mee&ng University of Virginia Mike Leipold

2 Two Different Approaches to Simultaneous Detec&on Fluorophores Elemental Tagging Fluorometer Induc&vely Coupled Plasma Mass Spectrometry (ICP- MS)

3 Two Different Approaches to Simultaneous Detec&on 8 Alexa Fluorophores Elemental Tagging intensity wavelength LSR II CyTOF

4 Overview 1. CyTOF the machine, pros and cons 2. Panel development background, desired signals 3. Experimental procedures an&bodies, reagents 4. Running samples example data - LSRII vs CyTOF - Flu 21, 211

5 Part 1 The CyTOF

6 Single Cell Analysis - CyTOF TM Machine Flow Interface to ICP- MS - Nebulizer vaporizes Flow - Argon Plasma atomizes and ionizes - Mass analyzer Figure courtesy O. Ornatsky

7 Single Cell Analysis - CyTOFTM Machine Published in: Dmitry R. Bandura; Vladimir I. Baranov; Olga I. Ornatsky; Alexei Antonov; Robert Kinach; Xudong Lou; Serguei Pavlov; Sergey Vorobiev; John E. Dick; Scob D. Tanner; Anal. Chem. 29, 81, DOI: 1.121/ac9149w Copyright 29 American Chemical Society

8 Display of Cell Events Monocyte? CD8+ T cell? B cell? - Push analogous to &me: 76,8 pushes/sec: 22 displayed here (1/4 of data)

9 Pros of CyTOF - Advantages: - Minimal "spectral" overlap higher dimensionality (more probes at once) - Quan&ta&ve broad dynamic range - Not light- or &me- sensi&ve - Minimal background - no analogy to autofluorescence: low/nil biological background for lanthanides

10 Cons of CyTOF - Disadvantages: - Destruc&ve: (currently) no way to recover interes&ng cells - Slower: limit of 1 cells/sec; prac&cal limit for best resolu&on oken in ~4 cells/sec range - Cell transmission efficiency: only ~2-3% of cells that enter machine get counted - Ion transmission efficiency: only ~1 in 1, ions that enter machine get counted

11 Cons of CyTOF - Disadvantages: - Postprocessing: cannot set on fly gates to get only live intact singlets - Events registered by CyTOF contains debris, doublets, etc - Might only get ~5% of total events as live intact singlets - No analogy to FSC or SSC: MUST have marker (M n+ ) for any ga&ng

12 Single Cell Analysis - CyTOF TM Machine - CyTOFv1 ~21- May Mass window ~93 units: eg, AW CyTOFv2 May 213- current - Mass window ~135 units: eg, AW More automated tuning, including recordkeeping - Improvements in ion op&cs - &ghter peaks, less "spillover"

13 Part 2 Panel Design

14 Relevant Issues to Panel Design 1. Background any non- desired signal - contamina&ng signals - an&body &ter (not discussed) 2. Desired Signal - an&body clone - metal label analogous to fluorophore choice

15 Relevant Issues to Panel Design Background Signals * Desired signal only at Mass "M" 1. M+16 oxide cannot eliminate, can only limit 2. Metal salt impuri&es M+1, M- 1, etc; Ln 3. Environmental contamina&on sample 4. Instrument * All poten&al sources of background, even spillover!

16 Relevant Issues to Panel Design Background Signals * Desired signal only at Mass "M" 1. M+16 oxide cannot eliminate, can only limit - Proper daily machine tuning - Lower AW Ln = worse

17 Daily Tuning Liquid Tuning Solu&on Current profile Make- up Gas profile

18 DVS Elemental Beads "Cells" - Polystyrene beads: contain nat. abund. Eu; or La, Pr, Tb, Tm, Lu - Act as "cells" run in acquisi&on mode (vs. tuning/solu&on mode) 1 4 (La139)Dd: (La139)Dd Makeup Gas Flow Rates - Median Dual counts vs AW 2 MG = (Pr141)Dd: (Pr141)Dd Median Dual Count 1 (Tb159)Dd: (Tb159)Dd Atomic Mass (Tm169)Dd: (Tm169)Dd

19 Effect of Tuning Beads - Oxida&on Linear Scale Median Dual counts Log Scale Median Dual counts Median Dual Count La139 Pr141 Tb159 Tm169 Lu175 "Gd155" "Gd157" "Re185" Median Dual Count La139 Pr141 Tb159 Tm169 Lu175 "Gd155" "Gd157" "Re185" * Makeup Gas Flow Rate (L/min) * Makeup Gas Flow Rate (L/min) - At high MG flow rate, oxida&on can be significant (decreases M, increases M+16 spillover) - Even a few percent can be significant for high- abundance markers (eg, CD57, histone proteins, CD45, etc)

20 Relevant Issues to Panel Design Background Signals * Desired signal only at Mass "M" 2. Metal salt impuri&es a) Few lanthanides are naturally 1% monoisotopic - e.g., Nd144 signal in Nd145 salt - M+1, M- 1, etc b) Other lanthanide contamina&ons all Ln chemistry is similar - La139 most common contaminant

21 Purity of Isotopes Liquid Stock Solu&ons CyTOF Channel Single- Isotope Sample - - M+1 impuri&es some isotopes worse than others M+16 oxida&on impuri&es affects all isotopes; lower AW worst

22 BD Ig Κ Capture Beads and MAXPAR An&bodies M M M+1 Sm # Cells # Cells 1 5 # Cells IgD(Nd146)Dd: IgD CD85j(Sm147)Dd: CD85j Nd148(Nd148)Dd: Nd148 No "Sm146" is Nd * Spillover due to elemental and/or isotopic impuri&es, not strictly mass proximity

23 Major Contaminants/Spillovers: Percentage of M - M+1 and M+16 are usually the major spillovers - Only Fluidigm metals shown; non- Fluidigm metals can be worse * Ir191 contam?

24 Major Contaminants/Spillovers: Cell Data * Formerly, Fluidigm sold Dy162 and Dy164, but not Dy163 Dy purity issues Regular Phenotyping panel: CD45RA- Dy162 CD2- Dy164 * no Dy163 an&body in experiment, so not usually monitored

25 Major Contaminants/Spillovers: Cell Data CD45RA- Dy162 CD2- Dy164 * no Dy163 an&body in experiment! Magenta: both Dy162, Dy164 Blue: Dy162 only Red: Dy164 only 1 CD3+ 1 B cells % of Max no CD2 % of Max high CD2, high CD45RA (Dy163)Dd (Dy163)Dd

26 Relevant Issues to Panel Design Background Signals 3. Environmental contamina&on - Reagents PFA lots with micropar&cles, MeOH with free metal, Ar gas with Sn - Biological sourcing Iodine (free I vs IdU), Pt in cispla&n- treated donor samples - Lab dust - Barium, lead, etc dish soap, syringes, reagent sourcing, striker flint, etc

27 Relevant Issues to Panel Design Background Signals 4. Instrument Abundance Sensi&vity lek (M- 1) and right (M+1) leg of ion peak - CyTOFv1: - 1% - CyTOFv2: <.3% 4 # Cells Worse with high AW masses: peak less Gaussian, M+1 leg wider (Sm147)Dd: CD85j

28 Major Contaminants/Spillovers: Percentage of M * CyTOF spillover - s&ll much less than Fluorescence spillover! * All propor&onal to signal at Mass "M" Match marker abundance with metal brightness - 1% of 3 is 3 (background) - 1% of 3 is 3 (probably not background) 2. Mutually exclusive lineages - T cell marker followed by B cell marker 3. Dump channels for most contaminated isotopes - analyze the "nega&ve" cells

29 Desired Signal Intensity- Choice of Metal Reminder: you cannot detect anything that doesn't have bound metal - No scaber to gate monocytes from lymphocytes, etc - Only about 3- fold difference in signal across lanthanides - Switching lanthanides might help only in borderline cases

30 Desired Signal Intensity- Choice of Metal 1 4 Lower sensitivity (139-15) # Cells CD56(Yb174)Dd: CD CD3(Nd15)Dd: CD CD16(Sm149)Dd: CD Higher sensitivity (165-17) # Cells CD56(Yb174)Dd CD3(Tm169)Dd CD16(Ho165)Dd

31 Desired Signal Intensity- Choice of Metal 1. "Dim" metals In, La, Pr, Nd, Sm - Bright/abundant markers (eg, CD57) - True bimodal (eg, CD27) 3 CD8+ 8 CD4+ 6 # Cells 2 # Cells (In113)Dd: CD CD27(Sm152)Dd: CD27

32 Desired Signal Intensity- Choice of Metal 2. "Bright" metals Tb, Dy, Tm, Yb - Dim/rare markers - "Smear" distribu&on (eg, CD45RA, CCR7) - Small fold- change TCRgd(Yb171)Dd: TCRgd14 TCRgd 3.29 # Cells CD CD3(Nd15)Dd: CD CD45RA(Dy162)Dd: CD45RA

33 Desired Signal Intensity- Choice of Metal 3. Recommend bivariates - one bright, one dim - two mediums "smear", or subpopula&ons CD2(Dy164)Dd: CD B cells 48.2 (Er166)Dd: CD CD14- CD CD14+ CD CD19(Nd142)Dd: CD (Sm154)Dd: CD14

34 Desired Signals CD85j as a Borderline Case CD4+ CD8+ Monocytes # Cells 2 1 CD85j CD85j+ 3.7 # Cells 1 5 CD85j CD85j # Cells 1 5 CD85j CD85j (Sm147)Dd: CD85j (Sm147)Dd: CD85j (Sm147)Dd: CD85j "nega&ve" subpopula&on "posi&ve"

35 Confirm New Markers With More Than One Donor TCRgd (Yb171)Dd: TCRgd TCRgd 3.8 (Yb171)Dd: TCRgd TCRgd (Nd15)Dd: CD (Nd15)Dd: CD3 1 4 CD14+ CD CD14+ CD CD33 (Er166)Dd: CD (Er166)Dd: CD CD14- CD CD14- CD (Sm154)Dd: CD (Sm154)Dd: CD14

36 Part 3 Experimental Procedure

37 CyTOF Sample Info 1. Reagent purity is paramount - MilliQ water only - trace metal pure salts and buffers, if possible 2. All samples - fixed and permeabilized - Thorough fixa&on - Fresh PFA! 3. MilliQ water washes at the end - Proper fixa&on ensure intact cells - Minimizes buffer salt introduc&on into machine 4. Titrate all reagents for your assay condi&ons - Commercial - Every batch of in- house reagents re- verify before use

38 CyTOF Sample Info Star&ng Cell Count * Remember: only 2-3% cell transmission efficiency - also, processing losses Cells Plated Live intact singlets Titration of cell numbers - Plated vs Live Intact Singlets 2K 28K 15, 5K 1M 76K 175K Live intact singlets 1, 5, R square Live intact Singlets.9895 Yao et al: down to 1K (major only) HIMC: recommend at least 5K 5, 1,, 1,5, Cell #, plated Slope Y-intercept when X=. X-intercept when Y=. Live intact Singlets.1829 ± ±

39 Metal Staining of Cells - DNA Intercalator - Labels any cell containing DNA regardless of whether any labelled an&bodies bind Schaferet al J.Organomet.Chem., 27, 692, Ornatsky et al, Anal. Chem., 28, 8,

40 Chela&ng Polymer Structures X8 linear polymer with ~22 chela&on sites DN3 branched dendrimer, ~16 chela&on sites

41 An&bodies X8 vs DN3 Comparison

42 Live- Dead maleimide- DOTA cispla&n

43 Normaliza&on EQ Four- Element Beads - Polystyrene beads: contain elemental Ce, Eu, Ho, Lu - burn like cells, but are defined composi&on

44 Normaliza&on Beads - Long Run&me Decrease in Signal Intensity 1:33 AM 6:55 PM

45 Normaliza&on Affects All Signals Noise amplifica&on! * Global Mean as baseline, rather than Minimum or Maximum - limits noise amplifica&on Normaliza&on CANNOT correct for improper machine cleaning/tuning (undetectable analyte signals). Finck et al, Cytometry A, 213

46 Signal Decrease Over Very Long Run- Times - Single file: >2 million cells = >3 hr 98% of start 73% of start * Highlights need for moving window normaliza&on Finck et al, Cytometry A, 213

47 Effects of Normaliza&on - Control sample split in half, run 8.5hr apart; ra&o aker manual ga&ng Freq Parent Median Intensity * Freq Parent fairly robust; only small gains from normaliza&on * Median Intensity more affected by normaliza&on (phosphoflow...)

48 Part 4 Sample Running

49 Running Samples 1. Warm up machine ~15-2 min to create and maintain fully hot and stable plasma 2. Tune machine maximize M signal while minimizing M+16 oxide forma&on 3. Finish washing cells final wash in MilliQ water, then resuspension and dilu&on in MilliQ water. - residual buffer salts can affect Current value 4. Resuspend and filter immediately before injec&on - Reduces aggregates (clogs) 5. Appropriate dilu&on minimize doublets - Around 1 cell event/screen refresh; usually ~75K/mL on cell counter

50 Standard CyTOF Output Appearance 15 - Spike at ~ is legi&mate ( true zero ) - Flow data is usually distributed above and below zero due to comps and auto- fluorescence. # Cells 1 5 CD CD * FlowJo se~ngs must be changed for CyTOF data CD3(Nd15)Dd: CD3

51 1 4 Intact cells DNA1(Ir191)Dd: DNA DNA1(Ir191)Dd: DNA Intact singlets DNA2(Ir193)Dd: DNA Cell_length: Cell_length CD33(Er166)Dd: CD Monocytes 3.7 Lymphocytes 69.1 DNA1(Ir191)Dd: DNA Live cells CD14(Sm154)Dd: CD Dead(In115)Dd: Dead

52 CyTOF vs LSRII Representa&ve Examples B Cells CD3+ Cells CD3- Cells Fluorescence <PE-Cy7-A>: CD <APC-Cy7-A>: CD CD CD <FITC-A>: CD NK cells <Am Cyan-A>: IgD <PerCP-Cy5-5-A>: CD <Pacific Blue-A>: CD16 Mass CD27(Sm152)Dd: CD CD8(Nd144)Dd: CD CD CD4+ 61 CD56(Yb174)Dd: CD NK cells IgD(Nd146)Dd: IgD CD4(Nd143)Dd: CD CD16(Sm149)Dd: CD16

53 Flu CyTOF Phenotyping Panel - Collapse 6 tube LSRII panel into 1 tube CyTOF panel 23 markers CD14 CD33 CD3 CD4 CD8 TCRgd CD19 CD2 CD16 CD56 CD45RA CCR7 CD25 CD127 CD27 CD28 CD38 CD85j IgD HLADR CD24 CD94 CD161

54 Flu 21, Flu 211 Studies - On- going Flu vaccine studies U19 - CyTOF Phenotyping 21, 211 years - Day (pre- vaccina&on) only Study 15, 17, 18 - Day, 1, 4, 7 Study : >3 unique donors between- donor variability Age Band (y/o) < >65 Total Male Female Total

55 21 Flu Studies Consistency 284 Control - Same Draw, Same Year Replicates

56 Flu 211, 212 Studies Consistency 885 Control - Generally good year- to- year agreement from same 885 donor draw Naive CD8+ IgD+ CD27- B cells 8 1 Percentage of Parent Percentage of Parent Men Men Year Year

57 Flu 211, 212 Studies Consistency 885 Control - Some&mes greater variability in less- frequent popula&ons CD4+ Treg 1 8 Percentage of Parent Men Year

58 Flu 21, Flu 211 Studies Donor Variability 8 **** *** B cells total **** **** Percentage of Parent Mann- Whitney two- tailed * <.5 ** <.1 *** <.1 **** <.1 Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65

59 Flu 21, Flu 211 Studies Donor Variability Mann- Whitney two- tailed * <.5 ** <.1 *** <.1 **** <.1 Percentage of Parent **** CD4+ total **** *** Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65 Percentage of Parent **** *** ** CD4+ Naive **** **** * Percentage of Parent CD4+ Effector Memory *** * * Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65 Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65

60 Flu 21, Flu 211 Studies Donor Variability Mann- Whitney two- tailed * <.5 ** <.1 *** <.1 **** <.1 Percentage of Parent * * CD8+ total **** *** * Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65 Percentage of Parent **** **** **** CD8+ Naive **** **** **** Percentage of Parent CD8+ Effector Memory **** **** * **** *** **** -2 Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65 Men <35 Men Men >65 Women <35 Gender and Age Band Women Women >65

61 Study (Sm152)Dd: CD CD27+ CD Day Day 1 (Sm152)Dd: CD CD27+ CD (Eu151)Dd: CD (Eu151)Dd: CD38 (Sm152)Dd: CD Day 4 Day 7 CD27+ CD (Sm152)Dd: CD CD27+ CD (Eu151)Dd: CD (Eu151)Dd: CD38

62 Conclusions 1. Proper tuning of the CyTOF ensures minimal sample oxida&on. 2. Metal purity (both elemental and isotopic) is highly important. 3. The choice of metal label is dependent upon marker abundance, modality, and resolu&on needed (fold- change). 4. All samples must be stained with metals to be detected by the CyTOF. Proper fixa&on is important, as are MilliQ water washes. 5. Head- to- head comparisons with LSRII assays are consistent. 6. The Flu studies are an example of how a CyTOF panel can be used to monitor the surface phenotype for hundreds of donors to detect varia&ons between gender, age- band, and &me- point.

63 Acknowledgements - HIMC staff - Henrik Mei, Meena Malipotlalla, Holden Maecker - External CyTOF: - Nolan lab: Sean Bendall, Erin Simonds, Eli Zunder, Bernd Bodenmiller - Davis lab: Evan Newell, Bill O'Gorman, Peber Brodin - Fluidigm/DVS Sciences - U19 NIH Grants and HIMC Service Center fees hbp://himc.stanford.edu

Categorical analysis of human T cell heterogeneity with One-SENSE

Categorical analysis of human T cell heterogeneity with One-SENSE 1 2 3 4 Supplementary Information Categorical analysis of human T cell heterogeneity with One-SENSE 5 6 Running title: T cell Analysis by One-SENSE 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 24 25 26

More information

Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry

Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry Nat Met, April 2014 Nat Med, April 2014 Next-Generation Immunohistochemistry: Multiplex tissue imaging with mass cytometry Journal Club Timo Böge Overview Introduction Conventional Immunohistochemistry

More information

Mass Cytometry Publication List

Mass Cytometry Publication List Mass Cytometry Publication List 2014 Publications 1 Becher, B., et al. High-dimensional analysis of the murine myeloid cell system. Nat Immunol 15 (12): 1181-1189, 2014. Behbehani, G.K., et al. Transient

More information

Do Your Flow Cytometric LDTs. Validation Guidelines? Fiona E. Craig, MD University of Pittsburgh School of Medicine

Do Your Flow Cytometric LDTs. Validation Guidelines? Fiona E. Craig, MD University of Pittsburgh School of Medicine Do Your Flow Cytometric LDTs Conform to the ICSH ICCS Validation Guidelines? Fiona E. Craig, MD University of Pittsburgh School of Medicine How should LDTs be validated? Accuracy Specificity Sensitivity

More information

DuraClone IM. Standardized phenotyping panels for studies of the human immune system

DuraClone IM. Standardized phenotyping panels for studies of the human immune system DuraClone IM Standardized phenotyping panels for studies of the human immune system Standardize with the experts, adopt DuraClone IM Tubes. Clinical research studies require accurate, reproducible results

More information

DURACLONE IF BE CERTAIN ABOUT THE RESPONSE. l res. a il n c n. For Research Use Only - Not for use in Diagnostic procedures

DURACLONE IF BE CERTAIN ABOUT THE RESPONSE. l res. a il n c n. For Research Use Only - Not for use in Diagnostic procedures DURACLONE IF earch tria l res lc a om ic il n c n nio pa Yo ur BE CERTAIN ABOUT THE RESPONSE For Research Use Only - Not for use in Diagnostic procedures BE CERTAIN ABOUT THE RESPONSE The sensitive and

More information

Fluorochrome Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 CTLA-4 CTLA-4 CD15 CD3 FITC. Bio) PD-1 (MIH4, BD) ICOS (C398.4A, Biolegend) PD-L1 (MIH1, BD)

Fluorochrome Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 CTLA-4 CTLA-4 CD15 CD3 FITC. Bio) PD-1 (MIH4, BD) ICOS (C398.4A, Biolegend) PD-L1 (MIH1, BD) Additional file : Table S. Antibodies used for panel stain to identify peripheral immune cell subsets. Panel : PD- signaling; Panel : CD + T cells, CD + T cells, B cells; Panel : Tregs; Panel :, -T, cdc,

More information

MHC MULTIMER PROFICIENCY PANEL 2017

MHC MULTIMER PROFICIENCY PANEL 2017 MHC MULTIMER PROFICIENCY PANEL 2017 August 2017 CONTACT Charlotte Halgreen ProficiencyPanel@immudex.com FOR MORE INFORMATION www.proficiencypanel.com MHC MULTIMER PROFICIENCY PANEL 2017 This report summarizes

More information

Optimizing Intracellular Flow Cytometry:

Optimizing Intracellular Flow Cytometry: Optimizing Intracellular Flow Cytometry: Simultaneous Detection of Cytokines and Transcription Factors Presented by Jurg Rohrer, PhD, BD Biosciences 23-10780-00 Outline Introduction Cytokines Transcription

More information

Gladstone Institutes, University of California (UCSF), San Francisco, USA

Gladstone Institutes, University of California (UCSF), San Francisco, USA Fluorescence-linked Antigen Quantification (FLAQ) Assay for Fast Quantification of HIV-1 p24 Gag Marianne Gesner, Mekhala Maiti, Robert Grant and Marielle Cavrois * Gladstone Institutes, University of

More information

Performance Characteristics of the Agilent High Matrix Sample Introduction (HMI) Accessory for the 7500 Series ICP-MS. Product Overview.

Performance Characteristics of the Agilent High Matrix Sample Introduction (HMI) Accessory for the 7500 Series ICP-MS. Product Overview. Performance Characteristics of the Agilent High Matrix Sample Introduction (HMI) Accessory for the 7500 Series ICP-MS Product Overview Introduction The determination of multiple trace elements in high-matrix

More information

DEVELOPMENTAL VALIDATION OF SPERM HY-LITER EXPRESS Jennifer Old, Chris Martersteck, Anna Kalinina, Independent Forensics, Lombard IL

DEVELOPMENTAL VALIDATION OF SPERM HY-LITER EXPRESS Jennifer Old, Chris Martersteck, Anna Kalinina, Independent Forensics, Lombard IL DEVELOPMENTAL VALIDATION OF SPERM HY-LITER EXPRESS Jennifer Old, Chris Martersteck, Anna Kalinina, Independent Forensics, Lombard IL Background and Introduction SPERM HY-LITER is the first immunofluorescent

More information

Superior Fluorescent Labeling Dyes Spanning the Full Visible Spectrum...1. Trademarks: HiLyte Fluor (AnaSpec, Inc.)

Superior Fluorescent Labeling Dyes Spanning the Full Visible Spectrum...1. Trademarks: HiLyte Fluor (AnaSpec, Inc.) Table of Contents Fluor TM Labeling Dyes Superior Fluorescent Labeling Dyes Spanning the Full Visible Spectrum....1 Fluor TM 405 Dye, an Excellent Alternative to Alexa Fluor 405 & DyLight 405....2 Fluor

More information

Mass Cytometry Applications for Immunology Research. Pub Note PN 13 01_150505

Mass Cytometry Applications for Immunology Research. Pub Note PN 13 01_150505 Mass Cytometry Applications for Immunology Research Pub Note PN 13 01_150505 Contents Continuum of CD8 + T cell phenotypes revealed by deep profiling with mass cytometry 3 The transcriptional landscape

More information

Supporting Information

Supporting Information Supporting Information Idoyaga et al. 10.1073/pnas.0812247106 SSC a) Single cell suspension 99 Aqua b) Live cells 96 -W c) Singlets 92 -A CD19+ER119 d) CD19 ER119 cells 97 CD3 e) CD3 cells 27 f) DX5 cells

More information

Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD-

Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- Supplementary Methods Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- L1 (10F.9G2, rat IgG2b, k), and PD-L2 (3.2, mouse IgG1) have been described (24). Anti-CTLA-4 (clone

More information

Analysis of Micronutrients in Milk by Flame Atomic Absorption Using FAST Flame Sample Automation for Increased Sample Throughput

Analysis of Micronutrients in Milk by Flame Atomic Absorption Using FAST Flame Sample Automation for Increased Sample Throughput APPLICATION NOTE Atomic Absorption Author: Nick Spivey PerkinElmer, Inc. Shelton, CT Analysis of Micronutrients in Milk by Flame Atomic Absorption Using FAST Flame Sample Automation for Increased Sample

More information

Optimizing Intracellular Flow Cytometry

Optimizing Intracellular Flow Cytometry Optimizing Intracellular Flow Cytometry Detection of Cytokines, Transcription Factors, and Phosphoprotein by Flow Cytometry Presented by Erika O Donnell, PhD, BD Biosciences 23-14876-00 Outline Basic principles

More information

Activity # 4. Measurement of Enzyme Activity

Activity # 4. Measurement of Enzyme Activity Activity # 4. Measurement of Enzyme Activity Adapted from Experimental Molecular Biology Lab Manual, Stephen H. Munroe, Marquette University Week 1 Protein Extraction, Concentration Measurement & Instrument

More information

of human hair and nails. Part I. Analytical methodology, Sci. Tot. Environ. 2000, 250/1-3,

of human hair and nails. Part I. Analytical methodology, Sci. Tot. Environ. 2000, 250/1-3, Reference data Biomonitoring Trace elements in human biological material References cited below: 1. Rodushkin I., Ödman F., Branth S. Multielement analysis of whole blood by high resolution inductively

More information

Phospholipid characterization by a TQ-MS data based identification scheme

Phospholipid characterization by a TQ-MS data based identification scheme P-CN1716E Phospholipid characterization by a TQ-MS data based identification scheme ASMS 2017 MP-406 Tsuyoshi Nakanishi 1, Masaki Yamada 1, Ningombam Sanjib Meitei 2, 3 1 Shimadzu Corporation, Kyoto, Japan,

More information

Understanding Poor Vaccine Responses: Transcriptomics of Vaccine Failure

Understanding Poor Vaccine Responses: Transcriptomics of Vaccine Failure Final Report - Summer Research Project 1 Introduction Understanding Poor Vaccine Responses: Transcriptomics of Vaccine Failure Katherine Miller, Year 3, Dalhousie University (Supervisor: Dr. Lisa Barrett,

More information

Evaluating the performance of the OneNeb Series 2 Nebulizer with PerkinElmer Optima 7/8x00 Series ICP-OES systems

Evaluating the performance of the OneNeb Series 2 Nebulizer with PerkinElmer Optima 7/8x00 Series ICP-OES systems Evaluating the performance of the OneNeb Series 2 Nebulizer with PerkinElmer Optima 7/8x Series ICP-OES systems Competitive Comparison Introduction The nebulizer is a novel nebulizer design that offers

More information

Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder I Nakaya, Kousik

Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder I Nakaya, Kousik SUPPLEMENTARY FIGURES 1-19 T H 2 response to cysteine-proteases requires dendritic cell-basophil cooperation via ROS mediated signaling Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder

More information

Business Statistics Probability

Business Statistics Probability Business Statistics The following was provided by Dr. Suzanne Delaney, and is a comprehensive review of Business Statistics. The workshop instructor will provide relevant examples during the Skills Assessment

More information

Comparison of Simultaneous Nitrofuran (NF) and Chloramphenicol (CAP) Analytical Methods for AOAC Validation Study

Comparison of Simultaneous Nitrofuran (NF) and Chloramphenicol (CAP) Analytical Methods for AOAC Validation Study Comparison of Simultaneous Nitrofuran (NF) and Chloramphenicol (CAP) Analytical Methods for AOAC Validation Study Haejung An Pacific Regional Lab-Southwest, FDA, Irvine, CA Three NF/CAP Methods Published

More information

ab65344 Uric Acid Assay Kit (Colorimetric/Fluorometric)

ab65344 Uric Acid Assay Kit (Colorimetric/Fluorometric) ab65344 Uric Acid Assay Kit (Colorimetric/Fluorometric) Instructions for Use For the rapid, sensitive and accurate measurement of uric acid in various samples. This product is for research use only and

More information

Documentation, Codebook, and Frequencies

Documentation, Codebook, and Frequencies Documentation, Codebook, and Frequencies Laboratory Component: Urinary Iodine Survey Years: 2005 to 2006 SAS Export File: UIO_D.XPT First Published: July 2008 Last revised: N/A NHANES 2005 2006 Data Documentation

More information

Naive, memory and regulatory T lymphocytes populations analysis

Naive, memory and regulatory T lymphocytes populations analysis Naive, memory and regulatory T lymphocytes populations analysis Jaen Olivier, PhD ojaen@beckmancoulter.com Cellular Analysis application specialist Beckman Coulter France Introduction Flow cytometric analysis

More information

QIAsymphony DSP Circulating DNA Kit

QIAsymphony DSP Circulating DNA Kit QIAsymphony DSP Circulating DNA Kit February 2017 Performance Characteristics 937556 Sample to Insight Contents Performance Characteristics... 4 Basic performance... 4 Run precision... 6 Equivalent performance

More information

BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit

BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit Instruction Manual Catalog No. 560485 ii Mouse Th1/Th2/Th17 Cytokine Kit 2009, Becton, Dickinson and Company. All rights reserved. No part

More information

Integrated Targeted Quantitation Method for Insulin and its Therapeutic Analogs

Integrated Targeted Quantitation Method for Insulin and its Therapeutic Analogs Integrated Targeted Quantitation Method for Insulin and its Therapeutic Analogs Eric Niederkofler, 1 Dobrin Nedelkov, 1 Urban Kiernan, 1 David Phillips, 1 Kemmons Tubbs, 1 Scott Peterman, 2 Bryan Krastins,

More information

ab CytoPainter Golgi/ER Staining Kit

ab CytoPainter Golgi/ER Staining Kit ab139485 CytoPainter Golgi/ER Staining Kit Instructions for Use Designed to detect Golgi bodies and endoplasmic reticulum by microscopy This product is for research use only and is not intended for diagnostic

More information

Supplementary Figure 1

Supplementary Figure 1 Supplementary Figure 1 FMO control (no HLA-DR antibody) FMO control (no HLA-DR antibody) Supplementary Figure 1. Gating strategy for HLA-DR+ T-cells. A gate was drawn around lymphocytes based on cell size

More information

MagniSort Mouse CD4 Naive T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures.

MagniSort Mouse CD4 Naive T cell Enrichment Kit Catalog Number: RUO: For Research Use Only. Not for use in diagnostic procedures. Page 1 of 2 MagniSort Mouse CD4 Naive T cell Enrichment Kit RUO: For Research Use Only. Not for use in diagnostic procedures. Mouse splenocytes were unsorted (left) or sorted with the MagniSort Mouse CD4

More information

CoQ10(Coenzyme Q10) ELISA Kit

CoQ10(Coenzyme Q10) ELISA Kit CoQ10(Coenzyme Q10) ELISA Kit Catalogue No.: EU0196 Size: 48T/96T Reactivity: Universal Detection Range: 0.781-50ng/ml Sensitivity:

More information

Low-Pressure Chromium Speciation in P2O5 using the Spectro ARCOS MV ICP and the ESI SC-DX chromfast System

Low-Pressure Chromium Speciation in P2O5 using the Spectro ARCOS MV ICP and the ESI SC-DX chromfast System Low-Pressure Chromium Speciation in P2O5 using the Spectro ARCOS MV ICP and the ESI SC-DX chromfast System Dion Tsourides AAFPCO Meeting Albuquerque, NM (Acknowledgements ESI) Introduction Chromium exists

More information

UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes

UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes Catalin E. Doneanu, Weibin Chen, and Jeffrey R. Mazzeo Waters Corporation, Milford, MA, U.S. A P P L I C AT ION B E N E F

More information

Application Note. Productivity and Efficiency of 6-Color BD Multitest and BD Trucount Technologies for Enumeration of Lymphocyte Subsets

Application Note. Productivity and Efficiency of 6-Color BD Multitest and BD Trucount Technologies for Enumeration of Lymphocyte Subsets Sept 27 Productivity and Efficiency of 6-Color BD Multitest and BD Trucount Technologies for Enumeration of Lymphocyte Subsets Phillip Ruiz, MD, PhD*, Michael J. Borowitz, MD, PhD**, Henok Tilahun***,

More information

PointCare NOW TM Technical Note

PointCare NOW TM Technical Note PointCare NOW TM Technical Note Introduction to our Technology And Summary of Evaluation and Performance data 1 of 19 Introduction The PointCare NOW is a portable, and very fast, diagnostic instrument

More information

Supplementary Figure 1. mtor LysM and Rictor LysM mice have normal cellularity and percentages of hematopoe>c cells. a. Cell numbers of lung, liver,

Supplementary Figure 1. mtor LysM and Rictor LysM mice have normal cellularity and percentages of hematopoe>c cells. a. Cell numbers of lung, liver, a. b. c. Supplementary Figure 1. mtor LysM and Rictor LysM mice have normal cellularity and percentages of hematopoe>c cells. a. Cell numbers of lung, liver, and spleen. b. Cell numbers of bone marrow

More information

Determination of Calcium in Serum Samples by AAS Using a Fuel Lean Flame

Determination of Calcium in Serum Samples by AAS Using a Fuel Lean Flame Determination of Calcium in Serum Samples by AAS Using a Fuel Lean Flame Application Note Atomic Absorption Authors Michael J. Sommer Michael G. Rutman Elizabeth Wask-Rotter Howard Wagoner and Edward T.

More information

A complete next-generation sequencing workfl ow for circulating cell-free DNA isolation and analysis

A complete next-generation sequencing workfl ow for circulating cell-free DNA isolation and analysis APPLICATION NOTE Cell-Free DNA Isolation Kit A complete next-generation sequencing workfl ow for circulating cell-free DNA isolation and analysis Abstract Circulating cell-free DNA (cfdna) has been shown

More information

SUPPORTING INFORMATIONS

SUPPORTING INFORMATIONS SUPPORTING INFORMATIONS Mice MT/ret RetCD3ε KO α-cd25 treated MT/ret Age 1 month 3 mnths 6 months 1 month 3 months 6 months 1 month 3 months 6 months 2/87 Survival 87/87 incidence of 17/87 1 ary tumor

More information

Kerrie Clerici, Michael Swain, Dominic Fernandez, Julia Schulz, Matthew Archer, Janine Campbell

Kerrie Clerici, Michael Swain, Dominic Fernandez, Julia Schulz, Matthew Archer, Janine Campbell Minimal Residual Disease (MRD) Testing by Flow Cytometry for childhood Precursor B Cell Acute Lymphoblastic Leukaemia Royal Children s Hospital experience. Kerrie Clerici, Michael Swain, Dominic Fernandez,

More information

Controlling cell-based bioassay performance through controlled preparation of bioassayready

Controlling cell-based bioassay performance through controlled preparation of bioassayready Controlling cell-based bioassay performance through controlled preparation of bioassayready cells Teresa Surowy September 2013 Promega Corporation Introduction The importance of cell-based bioassays and

More information

Challenges in Development and Validation of an Intracellular Cytokine Staining assay

Challenges in Development and Validation of an Intracellular Cytokine Staining assay Challenges in Development and Validation of an Intracellular Cytokine Staining assay Jenny Hendriks, Crucell Hatching @ EBF, Brussels, June 202 www.crucell.com Vaccines vs Protein therapeutics Protein

More information

ExoQuick PLUS Exosome Purification Kit for Serum & Plasma

ExoQuick PLUS Exosome Purification Kit for Serum & Plasma ExoQuick PLUS Exosome Purification Kit for Serum & Plasma Cat# EQPL10A-1 User Manual Store kit at +4 0 C Version 2 2/22/2017 A limited-use label license covers this product. By use of this product, you

More information

Lymphoma diagnosis on lymph node biopsies: classical diagnostic approach

Lymphoma diagnosis on lymph node biopsies: classical diagnostic approach overview Introduction Do we need tissue flowcytometry immunophenotyping in diagnosis and classification of NHL? Practical preanalytical & analytical considerations cases Lymphoma diagnosis on lymph node

More information

Supplemental Information. T Cells Enhance Autoimmunity by Restraining Regulatory T Cell Responses via an Interleukin-23-Dependent Mechanism

Supplemental Information. T Cells Enhance Autoimmunity by Restraining Regulatory T Cell Responses via an Interleukin-23-Dependent Mechanism Immunity, Volume 33 Supplemental Information T Cells Enhance Autoimmunity by Restraining Regulatory T Cell Responses via an Interleukin-23-Dependent Mechanism Franziska Petermann, Veit Rothhammer, Malte

More information

Annexin V-FITC Apoptosis Detection Kit with SYTOX

Annexin V-FITC Apoptosis Detection Kit with SYTOX ab14086 Annexin V-FITC Apoptosis Detection Kit with SYTOX Instructions for Use For the rapid, sensitive and accurate measurement of apoptosis in living cells. This product is for research use only and

More information

Comprehensive Forensic Toxicology Screening in Serum using On-Line SPE LC-MS/MS

Comprehensive Forensic Toxicology Screening in Serum using On-Line SPE LC-MS/MS Comprehensive Forensic Toxicology Screening in Serum using On-Line SPE LC-MS/MS SCIEX QTRAP 4500 LC-MS/MS System and Spark Holland PICO Adrian M. Taylor 1, Peter Ringeling 2, Martin Sibum 2, Stefan Sturm

More information

RayBio Annexin V-FITC Apoptosis Detection Kit Plus

RayBio Annexin V-FITC Apoptosis Detection Kit Plus RayBio Annexin V-FITC Apoptosis Detection Kit Plus User Manual Version 1.0 May 25, 2014 RayBio Annexin V-FITC Apoptosis (Cat#: 68FT-AnnVP-) RayBiotech, Inc. We Provide You With Excellent upport And ervice

More information

ab HRV 3C Protease Inhibitor Screening Kit (Colorimetric)

ab HRV 3C Protease Inhibitor Screening Kit (Colorimetric) Version 2 Last updated 2 March 2017 ab211089 HRV 3C Protease Inhibitor Screening Kit (Colorimetric) For the rapid, sensitive and accurate screening of potential HRV 3C Protease inhibitors. This product

More information

Mouse serum Insulin 200 tests

Mouse serum Insulin 200 tests Headquarters & Europe Office Cisbio Bioassays Phone: +33 (0)4 66 79 67 05 Fax: +33 (0)4 66 79 19 20 bioassays@cisbio.com cisbio_dd_pi_62in3pef-2µl USA Office Cisbio US, Inc. Phone: +1 888 963 4567 Fax:

More information

BioGraph Infiniti DynaMap Suite 3.0 for ProComp Infiniti March 2, SA7990B Ver 3.0

BioGraph Infiniti DynaMap Suite 3.0 for ProComp Infiniti March 2, SA7990B Ver 3.0 BioGraph Infiniti DynaMap Suite 3.0 for ProComp Infiniti March 2, 2006 SA7990B Ver 3.0 DynaMap Suite 3.0 for ProComp Infiniti Contents List P Inf 8 Ch EMG Bi-Lateral Difference.chs 3 P Inf 8 Ch EMG.chs

More information

Gentian Canine CRP Immunoassay Application Note for Abbott Architect * c4000

Gentian Canine CRP Immunoassay Application Note for Abbott Architect * c4000 v2-jan214 Gentian Canine CRP Immunoassay Application Note for Abbott Architect * c4 Intended Use The canine CRP immunoassay on Abbott's Architect c4 is an in vitro diagnostic test for quantitative determination

More information

ab Factor Xa Activity Assay Kit (Fluorometric)

ab Factor Xa Activity Assay Kit (Fluorometric) ab204711 Factor Xa Activity Assay Kit (Fluorometric) Instructions for Use For rapid, sensitive and accurate detection of Factor Xa activity. This product is for research use only and is not intended for

More information

Branched Chain Amino Acid (Leu, Ile, Val) Assay Kit

Branched Chain Amino Acid (Leu, Ile, Val) Assay Kit Branched Chain Amino Acid (Leu, Ile, Val) Assay Kit Catalog Number KA0824 100 assays Version: 03 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General

More information

Bead Based Assays for Cytokine Detection

Bead Based Assays for Cytokine Detection Bead Based Assays for Cytokine Detection September 27, 2014 6 th EFIS-EJI South East European Immunology School SEEIS 2014 Timisoara, Romania The Cells of the Immune System The Immune Reaction (Th2) (Th1)

More information

REDOX PROTEOMICS. Roman Zubarev.

REDOX PROTEOMICS. Roman Zubarev. REDOX PROTEOMICS Roman Zubarev Roman.Zubarev@ki.se Physiological Chemistry I, Department for Medical Biochemistry & Biophysics, Karolinska Institutet, Stockholm What is (RedOx) Proteomics? Proteomics -

More information

Successful flow cytometric immunophenotyping of body fluid specimens

Successful flow cytometric immunophenotyping of body fluid specimens Successful flow cytometric immunophenotyping of body fluid specimens Fiona E. Craig, MD Division of Hematopathology Mayo Clinic Arizona 2017 MFMER slide-1 Financial disclosure No conflicts 2017 MFMER slide-2

More information

Quantitative Analysis of Vit D Metabolites in Human Plasma using Exactive System

Quantitative Analysis of Vit D Metabolites in Human Plasma using Exactive System Quantitative Analysis of Vit D Metabolites in Human Plasma using Exactive System Marta Kozak Clinical Research Applications Group Thermo Fisher Scientific San Jose CA Clinical Research use only, Not for

More information

Elemental Scientific Semiconductor Catalog for Thermo ELEMENT2

Elemental Scientific Semiconductor Catalog for Thermo ELEMENT2 Elemental Scientific Semiconductor Catalog for Thermo ELEMENT2 Ordering Information You may contact us via phone, fax, e-mail, or through our website. EPOND S.A. CP 389 CH-1800 Vevey Switzerland Phone:

More information

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay Catalog # Description 172-5080 SingleShot Cell Lysis Kit, 100 x 50 µl reactions 172-5081 SingleShot Cell Lysis Kit, 500 x 50 µl reactions For research purposes only. Introduction The SingleShot Cell Lysis

More information

A Definitive Lipidomics Workflow for Human Plasma Utilizing Off-line Enrichment and Class Specific Separation of Phospholipids

A Definitive Lipidomics Workflow for Human Plasma Utilizing Off-line Enrichment and Class Specific Separation of Phospholipids A Definitive Lipidomics Workflow for Human Plasma Utilizing Off-line Enrichment and Class Specific Separation of Phospholipids Jeremy Netto, 1 Stephen Wong, 1 Federico Torta, 2 Pradeep Narayanaswamy, 2

More information

RayBio Human Phospho-DDR2 (Tyr740) and Total DDR2 ELISA Kit

RayBio Human Phospho-DDR2 (Tyr740) and Total DDR2 ELISA Kit RayBio Human Phospho-DDR2 (Tyr740) and Total DDR2 ELISA Kit Catalog #: PEL-DDR2-Y740-T User Manual Last revised March 22, 2018 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607

More information

7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit

7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit 7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit Item No. 600120 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS

More information

Manual. Transfer step. Figure 1. Workflow comparison of manual phenylalanine assay and automated GSP Neonatal Phenylalanine assay

Manual. Transfer step. Figure 1. Workflow comparison of manual phenylalanine assay and automated GSP Neonatal Phenylalanine assay TECHNICAL NOTE GSP Neonatal Phenylalanine kit 3308-0010/3308-001B GSP Neonatal Phenylalanine kit (3308-0010/3308-001B) is a fully Introduction automated enzymatic assay for the GSP system. Optimized for

More information

Mouse Th1/Th2 Cytokine CBA

Mouse Th1/Th2 Cytokine CBA BD Cytometric Bead Array Mouse Th1/Th2 Cytokine CBA Cat. No. 551287 BD Biosciences For research use only. Not for use in diagnostic or therapeutic procedures. Purchase does not include or carry any right

More information

Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary Materials and Methods

Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary Materials and Methods Silva et al. PTEN posttranslational inactivation and hyperactivation of the PI3K/Akt pathway sustain primary T cell leukemia viability Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary

More information

TOP-DOWN DETERMINATION OF SERUM INSULIN-LIKE GROWTH FACTORS BY LC-MS

TOP-DOWN DETERMINATION OF SERUM INSULIN-LIKE GROWTH FACTORS BY LC-MS TOP-DOWN DETERMINATION OF SERUM INSULIN-LIKE GROWTH FACTORS BY LC-MS Richard KT Kam, Ph.D. Scientific Officer, Department of Chemical Pathology Prince of Wales Hospital Prince of Wales Hospital, Hong Kong

More information

Exo-Glow TM Exosome Labeling Kits

Exo-Glow TM Exosome Labeling Kits Exo-Glow TM Exosome Labeling Kits Cat# EXOR100A-1 Cat# EXOG200A-1 Cat# EXOC300A-1 User Manual Store kit at -20 o C on receipt Version 8 3/10/2017 A limited-use label license covers this product. By use

More information

Serafino et al. Thymosin α1 activates complement receptor-mediated phagocytosis in human monocyte-derived macrophages. SUPPLEMENTARY FIGURES

Serafino et al. Thymosin α1 activates complement receptor-mediated phagocytosis in human monocyte-derived macrophages. SUPPLEMENTARY FIGURES Supplementary Fig. S1. Evaluation of the purity and maturation of macrophage cultures tested by flow cytometry. The lymphocytic/monocytic cellular fraction was isolated from buffy coats of healthy donors

More information

ab65656 Ascorbic Acid Assay Kit (Colorimetric)

ab65656 Ascorbic Acid Assay Kit (Colorimetric) ab65656 Ascorbic Acid Assay Kit (Colorimetric) Instructions for Use For the rapid, sensitive and accurate measurement of Ascorbic Acid in various biological samples. This product is for research use only

More information

RayBio DPP4 Inhibitor Screening Kit

RayBio DPP4 Inhibitor Screening Kit RayBio DPP4 Inhibitor Screening Kit User Manual Version 1.0 Mar 25, 2013 RayBio DPP4 Inhibitor Screening (Cat#: 68SR-DPP4-S100) RayBiotech, Inc. We Provide You With Excellent Support And Service Tel:(Toll

More information

Agilent 1260 Infinity Analytical SFC System

Agilent 1260 Infinity Analytical SFC System Agilent 1260 Infinity Analytical SFC System Installation Instructions: Checkout Procedure This technical note describes the checkout procedures for the Agilent 1260 Infinity Analytical SFC System. Contents

More information

CORESTA Recommended Method No. 84

CORESTA Recommended Method No. 84 Cooperation Centre for Scientific Research Relative to Tobacco E-Vapour Sub-Group CORESTA Recommended Method No. 84 DETERMINATION OF GLYCERIN, PROPYLENE GLYCOL, WATER, AND NICOTINE IN THE AEROSOL OF E-CIGARETTES

More information

LC/MS/MS SOLUTIONS FOR LIPIDOMICS. Biomarker and Omics Solutions FOR DISCOVERY AND TARGETED LIPIDOMICS

LC/MS/MS SOLUTIONS FOR LIPIDOMICS. Biomarker and Omics Solutions FOR DISCOVERY AND TARGETED LIPIDOMICS LC/MS/MS SOLUTIONS FOR LIPIDOMICS Biomarker and Omics Solutions FOR DISCOVERY AND TARGETED LIPIDOMICS Lipids play a key role in many biological processes, such as the formation of cell membranes and signaling

More information

RayBio Human Phosphotyrosine BTK ELISA Kit

RayBio Human Phosphotyrosine BTK ELISA Kit RayBio Human Phosphotyrosine BTK ELISA Kit Catalog #: PEL-BTK-Y User Manual Last revised August 10, 2016 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607 Parkway Lane, Suite

More information

RayBio Human ENA-78 ELISA Kit

RayBio Human ENA-78 ELISA Kit RayBio Human ENA-78 ELISA Kit Catalog #: ELH-ENA78 User Manual Last revised April 15, 2016 Caution: Extraordinarily useful information enclosed ISO 13485 Certified 3607 Parkway Lane, Suite 100 Norcross,

More information

BD Multitest IMK Kit

BD Multitest IMK Kit BD Multitest IMK Kit 50 Tests Catalog No. 340503 50 Tests with BD Trucount Tubes Catalog No. 340504 IVD 2016 BD. BD, the BD Logo and all other trademarks are property of Becton, Dickinson and Company.

More information

Utilizing AlphaLISA Technology to Screen for Inhibitors of the CTLA-4 Immune Checkpoint

Utilizing AlphaLISA Technology to Screen for Inhibitors of the CTLA-4 Immune Checkpoint APPLICATION NOTE AlphaLISA Technology Authors: Matthew Marunde Stephen Hurt PerkinElmer, Inc. Hopkinton, MA Utilizing AlphaLISA Technology to Screen for Inhibitors of the CTLA-4 Immune Checkpoint Introduction

More information

ab Lysosome/Cytotoxicity Dual Staining Kit

ab Lysosome/Cytotoxicity Dual Staining Kit ab133078 Lysosome/Cytotoxicity Dual Staining Kit Instructions for Use For studying lysosome function at the cellular level. This product is for research use only and is not intended for diagnostic use.

More information

Protocols in Cardiac CT Dr. Bruce Precious Dalhousie University Friday, April 15, 2016

Protocols in Cardiac CT Dr. Bruce Precious Dalhousie University Friday, April 15, 2016 Protocols in Cardiac CT Dr. Bruce Precious Dalhousie University Friday, April 15, 2016 Disclosure Statement: No Conflict of Interest I do not have an affiliation, financial or otherwise, with a pharmaceutical

More information

ab HMG-CoA Reductase Activity Assay Kit (Colorimetric)

ab HMG-CoA Reductase Activity Assay Kit (Colorimetric) ab204701 HMG-CoA Reductase Activity Assay Kit (Colorimetric) Instructions for Use For rapid, sensitive and accurate detection of HMG-CoA Reductase Activity. This product is for research use only and is

More information

Fizzy Extraction of Volatile and Semivolatile Compounds into the Gas Phase

Fizzy Extraction of Volatile and Semivolatile Compounds into the Gas Phase SUPPORTING INFORMATION Fizzy Extraction of Volatile and Semivolatile Compounds into the Gas Phase Cheng-Hao Chang, Pawel L. Urban* Department of Applied Chemistry, National Chiao Tung University 1001 University

More information

UCLA. Dynamics of Clonal Evolution: Immune System on a Chip. Caltech. Jim Heath, Caltech Chemistry

UCLA. Dynamics of Clonal Evolution: Immune System on a Chip. Caltech. Jim Heath, Caltech Chemistry Dynamics of Clonal Evolution: Immune System on a Chip Jim Heath, Caltech Chemistry Caltech David Baltimore Lab: Jonathan Tsai Heath Lab: Alex Balazs Chao Ma Lili Yang Ann Cheung Michael Bethune Gabe Kwong

More information

The Annexin V Apoptosis Assay

The Annexin V Apoptosis Assay The Annexin V Apoptosis Assay Development of the Annexin V Apoptosis Assay: 1990 Andree at al. found that a protein, Vascular Anticoagulant α, bound to phospholipid bilayers in a calcium dependent manner.

More information

METHOD DEVELOPMENT AND VALIDATION: QUANTIFICATION OF METALS IN LIQUIDS AND AEROSOL OF E-CIGARETTES

METHOD DEVELOPMENT AND VALIDATION: QUANTIFICATION OF METALS IN LIQUIDS AND AEROSOL OF E-CIGARETTES METHOD DEVELOPMENT AND VALIDATION: QUANTIFICATION OF METALS IN LIQUIDS AND AEROSOL OF E-CIGARETTES Sandra Otte, Sonja Nowak & Michael Intorp Imperial Tobacco Group; Special Analytes Laboratory Hamburg,

More information

AlphaScreen : A Straightforward and Powerful Alternative to ELISA. Martina Bielefeld-Sévigny Ph.D., R&D Director

AlphaScreen : A Straightforward and Powerful Alternative to ELISA. Martina Bielefeld-Sévigny Ph.D., R&D Director AlphaScreen : A Straightforward and Powerful Alternative to ELISA Martina Bielefeld-Sévigny Ph.D., R&D Director Overview AlphaScreen - an alternative to ELISA Why an alternative to ELISA? Assay principle

More information

Clinical question. Screening tube. Diagnostic panel MRD. Clinical question

Clinical question. Screening tube. Diagnostic panel MRD. Clinical question OW CYTOMETRY UPDATES IN LYMPHOPROLIFERATIVE DISORDERS CANCER RESEARCH CENTER IBSAL UNIVERSITY & UNIVERSITY HOSPITAL, SALAMANCA (SPAIN) DISCLOSURES The EuroFlow Scientific Consortium Iamco-chairof receives

More information

RP-HPLC Analysis of Temozolomide in Pharmaceutical Dosage Forms

RP-HPLC Analysis of Temozolomide in Pharmaceutical Dosage Forms Asian Journal of Chemistry Vol. 22, No. 7 (2010), 5067-5071 RP-HPLC Analysis of Temozolomide in Pharmaceutical Dosage Forms A. LAKSHMANA RAO*, G. TARAKA RAMESH and J.V.L.N.S. RAO Department of Pharmaceutical

More information

Determination of metals in industrial wastewaters by microwave plasmaatomic emission spectrometry

Determination of metals in industrial wastewaters by microwave plasmaatomic emission spectrometry Determination of metals in industrial wastewaters by microwave plasmaatomic emission spectrometry Application note Environmental Author Terrance Hettipathirana Agilent Technologies Melbourne, Australia

More information

Copy Number Varia/on Detec/on. Alex Mawla UCD Genome Center Bioinforma5cs Core Tuesday June 16, 2015

Copy Number Varia/on Detec/on. Alex Mawla UCD Genome Center Bioinforma5cs Core Tuesday June 16, 2015 Copy Number Varia/on Detec/on Alex Mawla UCD Genome Center Bioinforma5cs Core Tuesday June 16, 2015 Today s Goals Understand the applica5on and capabili5es of using targe5ng sequencing and CNV calling

More information

STAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit

STAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit STAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit Catalog Number KA2176 96 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Principle of the Assay...

More information

Why use Signal-To-Noise as a Measure of MS Performance When it is Often Meaningless?

Why use Signal-To-Noise as a Measure of MS Performance When it is Often Meaningless? Why use Signal-To-Noise as a Measure of MS Performance When it is Often Meaningless? Technical Overview Authors Greg Wells, Harry Prest, and Charles William Russ IV, Agilent Technologies, Inc. 285 Centerville

More information

Human Alpha 1 microglobulin ELISA Kit

Human Alpha 1 microglobulin ELISA Kit Human Alpha 1 microglobulin ELISA Kit Catalogue No.: EH4144 Size: 48T/96T Reactivity: Human Range:0.625-40ng/ml Sensitivity:

More information

Rapid Analysis of Water-Soluble Vitamins in Infant Formula by Standard-Addition

Rapid Analysis of Water-Soluble Vitamins in Infant Formula by Standard-Addition Rapid Analysis of Water-Soluble Vitamins in Infant Formula by Standard-Addition Evelyn Goh Waters Pacific, Singapore APPLICATION BENEFITS This method allows for the simultaneous analysis of 12 water-soluble

More information

Method Comparison Report Semi-Annual 1/5/2018

Method Comparison Report Semi-Annual 1/5/2018 Method Comparison Report Semi-Annual 1/5/2018 Prepared for Carl Commissioner Regularatory Commission 123 Commission Drive Anytown, XX, 12345 Prepared by Dr. Mark Mainstay Clinical Laboratory Kennett Community

More information