Recommended Protocols for Phospho Protein Detection in Human Cells
|
|
|
- Amie Norton
- 6 years ago
- Views:
Transcription
1 Recommended Protocols for Phospho Protein Detection in Human Cells Depending on subcellular localization of the phospho protein of interest, as well as epitope susceptibility to cell fixing and permeabilizing agents, we recommend different protocols for our antibodies. For phenotyping of cell populations in multi color experiments, please use the table on pages 27 30, showing our human cell surface markers tested under different fixation and permeabilization conditions. Protocol I II III Fix buffers used for PBMCs: BD Phosflow Fix Buffer I BD Cytofix Buffer BD Cytofix Buffer (557870) (554655) (554655) Fix buffers used for Whole Blood: BD Phosflow Lyse/Fix BD Phosflow Lyse/Fix BD Phosflow Lyse/Fix Buffer (558049) Buffer (558049) Buffer (558049) Perm buffers used: BD Phosflow Perm/Wash I BD Phosflow Perm II BD Phosflow Perm III Buffer (557885) Buffer (558052) Buffer (558050) Specificity Clone Fluorochrome ERK 1/2 (pt202/py204) 20a LCK (py505) 4/Lck-Y505 p38 MAPK (pt180/py182) 36 ZAP70 (py319)/syk (py352) 17a Alexa C-Cbl (py700) 47/c-Cbl (Y700) Alexa Stat1 (py701) 4a Alexa /- + PE +/- + + Stat3 (py705) 4 Alexa /- + Stat3 (py727) 49 Alexa 488??? Alexa 647??? Stat4 (py693) 38/p-Stat4 Alexa Alexa Stat5 (py694) 47 Alexa /- + Stat6 (py641) 18 Alexa /- + /- 26
2 Validated Human Cell Surface Markers for Analysis Many cell surface markers are sensitive to fixation/permeabilization procedures and lose recognition by relevant antibodies afterwards. For your convenience, many antibodies conjugated to various fluorochromes have been tested under several conditions. The results are summarized below. Protocol I II III Fix buffers used for PBMCs: BD Phosflow Fix Buffer I BD Cytofix Buffer BD Cytofix Buffer (557870) (554655) (554655) Fix buffers used for Whole Blood: BD Phosflow Lyse/Fix BD Phosflow Lyse/Fix BD Phosflow Lyse/Fix Buffer (558049) Buffer (558049) Buffer (558049) Perm buffers used: BD Phosflow Perm/Wash I BD Phosflow Perm II BD Phosflow Perm III Buffer (557885) Buffer (558052) Buffer (558050) Specificity Clone Fluorochrome CD3 SK7 PerCP + + +/- PE-Cy /- APC-Cy7 + CD3 UCHT1 FITC Alexa CD3 HIT3a FITC - - CD4 L200 FITC PerCP PerCPCy CD4 RPA-T4 FITC CD4 SK3 PerCP PE-Cy CD5 UCHT2 CD8 HIT8a FITC Chapter 1 27
3 Specificity Clone Fluorochrome I II III CD8 RPA-T8 FITC PE-Cy APC-Cy7 + Alexa CD8 SK1 PerCP + +/- CD11a/LFA-1 G43-25B PE + +/- - CD11a/LFA-1 Hi111 PE + +/ CD11b ICRF44 PE + +/- +/- - - PE-Cy7 + - Alexa CD11c BLy6 PE + +/- +/- +/- +/- CD13 WM-15 PE +/- +/- CD14 M5E2 FITC + +/- - PE*** + +/- - PerCP-Cy5.5 + PE-Cy7 + - Alexa 488*** + +/- CD14 MoP9 FITC*** - PE*** CD15 W6D3 PE + + CD15 H198 PE + + CD16 3G8 PE + +/- - FITC +/- +/- PE-Cy Alexa 488 +/- CD16 B73.1 PE*** + +/- - CD16 NKP15 FITC - - CD CD19* HIB19 PE*** + +/- - FITC +/- +/- PerCP-Cy5.5 +/- PE-Cy7 - Alexa 488*** - Alexa 647*** +/- CD19 SJ25C1 PE*** + +/- - PE-Cy7 +/- - - CD20 2H7 FITC +/- - - PE CD20 L
4 Specificity Clone Fluorochrome I II III CD20 (I/C) H1(FB1) CD23 M-L233 PE + +/- - CD25 M-A251 FITC + + CD27 L128 FITC CD27 MT271 - CD28 CD28.2 FITC - - CD31 WM59 PE + +/- +/- CD33 WM53 PE + +/- +/- CD33 p67.6 PE +/- +/- CD34 8G12 PE + +/- - CD CD38 HB7 FITC CD38 HIT2 - CD40 5C3 FITC CD PE + + CD44 G44-26 PE CD45 HI30 CD45RA 5H9 FITC + + CD45RA HI100 FITC CD45RO UCHL1 FITC CD54 HA58 PE +/- +/- +/- CD56 B159 - FITC - PE-Cy7 - - Alexa Alexa CD56 MY31 PE +/- +/- CD56 NCAM CD69 FN50 FITC - - PE +/ CD79a (I/C) HM47 CD80 L307.4 PE DC+ DC+ Chapter 1 29
5 Specificity Clone Fluorochrome I II III CD83 HB15e PE + +/- +/- CD CD86 IT2.2 CD94 HP-3D9 FITC CD138 Mi15 CD195 2D7 FITC PE-Cy7 +/- - - CD195 3A9 - DC-SIGN DCN46 CCR7 - HLA-DR G46-6 FITC + + +/- PE + + * HLA-DR Tu36 PE + +/- - IgD IA6-2 - IgM G PE + +/- +/- Legend: DC Dendritic Cells + Positive - Negative +/- Inconclusive Stimulators used for Multi-Color Analysis of Human Whole Blood or PBMCs PMA IFNα IL-2 IL-4 IL-6 CD3/CD28 Activation perk 1/2 pp38 MAPK pstat1 pstat1 (+/-) pstat3 pstat3 pstat5 pstat5 pstat6 pzap70 plck Bulk Conc. Final Conc. For PBMC Final Conc. For Whole Blood PMA 1mg/ml 40nM 400nM IFNα 0.2mg/ml 10,000U/ml 40,000U/ml ( U/ml) IL µg/µl 0.1µg/ml IL-4 0.1µg/µl 0.1µg/ml IL-6 0.2µg/µl 0.1µg/ml 30
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
BD Flow Cytometry Reagents Multicolor Panels Designed for Optimal Resolution with the BD LSRFortessa X-20 Cell Analyzer
Multicolor Panels Designed for Optimal Resolution with the BD LSRFortessa X-2 Cell Analyzer Proper multicolor panel design takes into account fluorochrome brightness, antigen density, co-expression, and
Optimizing Intracellular Flow Cytometry:
Optimizing Intracellular Flow Cytometry: Simultaneous Detection of Cytokines and Transcription Factors An encore presentation by Jurg Rohrer, PhD, BD Biosciences 10.26.10 Outline Introduction Cytokines
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,
20 ml (1 ea) 40 ml (1 ea) 25 ml (1 ea) 7.5 ml (1 ea)
Technical Data Sheet Transcription Factor Phospho Buffer Set Product Information Material Number: 565575 25 Tests Component: 51-9011474 Description: 0.75X TFP Diluent Buffer 20 ml (1 ea) BD Pharmingen
Glow. Save. Let it. and. Get a Discount of 10% BD Biosciences Year End Promotion on a huge Selection of Reagent Solutions
Let it Glow and Save BD Biosciences Year End Promotion 2014 Get a Discount of 10% on a huge Selection of Reagent Solutions All fluorochrome labeled antibodies The complete BD CBA portfolio Kits and reagents
Application Information Bulletin: Human NK Cells Phenotypic characterizing of human Natural Killer (NK) cell populations in peripheral blood
Application Information Bulletin: Human NK Cells Phenotypic characterizing of human Natural Killer (NK) cell populations in peripheral blood Christopher A Fraker, Ph.D., University of Miami - Miami, Florida
MAIT cell function is modulated by PD-1 signaling in patients with active
MAIT cell function is modulated by PD-1 signaling in patients with active tuberculosis Jing Jiang, M.D., Xinjing Wang, M.D., Hongjuan An, M.Sc., Bingfen Yang, Ph.D., Zhihong Cao, M.Sc., Yanhua Liu, Ph.D.,
Supplementary Table e-1. Flow cytometry reagents and staining combinations
Supplementary data Supplementary Table e-1. Flow cytometry reagents and staining combinations Reagents Antibody Fluorochrome Clone Source conjugation CD3 FITC UCHT1 BD Biosciences CD3 PerCP-Cy5.5 SK7 Biolegend
BD Pharmingen. Human Th1/Th2/Th17 Phenotyping Kit. Technical Data Sheet. Product Information. Description Components:
Technical Data Sheet Human Th1/Th2/Th17 Phenotyping Kit Product Information Material Number: Size: Description Components: 560751 50 Tests BD Pharmingen 51-9006615 Human Th1/Th2/Th17 Phenotyping Cocktail
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
Supplementary Data. Treg phenotype
Supplementary Data Additional Experiment An additional experiment was performed using cryopreserved peripheral blood mononuclear cells (PBMC) derived from five renal cell carcinoma (RCC) patients [see
Supplementalgfigureg1gSchematicgdiagramgofgtumor1modellingg
SChinjectionh F:LuchLCLsh IVhinjectionh T:cellsh Monitorhforhtumorh growthhandhxeno: reactivehgvhd GVLgexperimentg kcbgvsgpbgt1cellse Xeno1reactiveg experimentg kcbgvsgpbgt1cellse IVhinjectionh 5xh,N^6
Online supplement. Phenotypic, functional and plasticity features of classical and alternatively activated
Online supplement Phenotypic, functional and plasticity features of classical and alternatively activated human macrophages Abdullah Al Tarique*, Jayden Logan *, Emma Thomas *, Patrick G Holt *, Peter
MACS Marker Screen. MACS Marker Screen. Catalog antibodies. antibodies CD4 (VIT4) CD11b. MACS Marker Screen antibodies. Catalog. antibodies CD2 CD14
MACS Marker Screen The performance of MACS Marker Screen was tested and compared with catalog reagents offered in single vials as liquid antibody reagents. The were reconstituted using 25 µl of deionized
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
Detailed step-by-step operating procedures for NK cell and CTL degranulation assays
Supplemental methods Detailed step-by-step operating procedures for NK cell and CTL degranulation assays Materials PBMC isolated from patients, relatives and healthy donors as control K562 cells (ATCC,
BD Phosflow T Cell Activation Kit
BD Phosflow T Cell Activation Kit Instruction Manual Catalog No. 560750 ii BD Phosflow T Cell Activation Kit The information in this guide is subject to change without notice. BD Biosciences reserves the
BD IMag Cell Separation System
BD IMag Cell Separation System Table of Contents Magnetic Cell Separation with BD IMag Particles.............................................................. 3 BD IMag Enrichment Protocol Flow Chart......................................................................
Immunobiology Handbook
Immunobiology Handbook Immunobiology Handbook BioLegend is ISO 9001:2008 and ISO 13485:2003 Certified Toll-Free Tel: (US & Canada): 1.877.BIOLEGEND (246.5343) Tel: 858.768.5800 World-Class Quality Superior
IL-6Rα IL-6RαT-KO KO. IL-6Rα f/f bp. f/f 628 bp deleted 368 bp. 500 bp
STD H 2 O WT KO IL-6Rα f/f IL-6Rα IL-6RαT-KO KO 1000 bp 500 bp f/f 628 bp deleted 368 bp Supplementary Figure 1 Confirmation of T-cell IL-6Rα deficiency. (a) Representative histograms and (b) quantification
Application Information Bulletin: Cell Signaling
Application Information Bulletin: Cell Signaling Detection of Both Surface Proteins and Intracellular Phospho-Determinants in Whole Blood Using the PerFix-p Reagent System William L. Godfrey 1, Li Yang
CLINICAL USE OF CELLULAR SUBPOPULATION ANALYSIS IN BM
CLINICAL USE OF CELLULAR SUBPOPULATION ANALYSIS IN BM CANCER RESEARCH CENTRE, UNIVERSITY AND UNIVERSITY HOSPITAL OF SALAMANCA (SPAIN)( Sao Paulo, 18th of April, 2009 IDENTIFICATION OF HPC (I) 1.- In vivo
Supplementary Materials
Supplementary Materials 43 Figure S1. CD123 in acute lymphoblastic leukemia and leukemia-initiating cells. A. CD123 (histograms) is highly and homogenously expressed in B-ALL blasts (as defined by live,
Complete Brilliance. Brilliant Violet. Novel Fluorophore Conjugates for Flow Cytometry. for the Violet Laser. Antibody Conjugates
Brilliant Violet Antibody Conjugates Complete Brilliance for the Violet Laser Novel Fluorophore Conjugates for Flow Cytometry Toll-Free Tel: (US & Canada): 1.877.BIOLEGEND (246.5343) Tel: 858.768.58 biolegend.com
Peptide stimulation and Intracellular Cytokine Staining
v Peptide stimulation and Intracellular Cytokine Staining Authors A. Cosma, S. Allgayer MATERIALS Date 01-02-2007 REAGENTS: Version 1.0 - PBMCs - Culture medium - Costimulating antibodies - Peptide pools
Supplemental Information. Human Circulating PD-1 + CXCR3 CXCR5 + Memory. Tfh Cells Are Highly Functional and Correlate
Immunity, Volume 39 Supplemental Information Human Circulating PD-1 + CXCR3 CXCR5 + Memory Tfh Cells Are Highly Functional and Correlate with Broadly Neutralizing HIV Antibody Responses Michela Locci,
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
Cover Page. The handle holds various files of this Leiden University dissertation.
Cover Page The handle http://hdl.handle.net/1887/23854 holds various files of this Leiden University dissertation. Author: Marel, Sander van der Title: Gene and cell therapy based treatment strategies
Supplementary Figure 1. Immune profiles of untreated and PD-1 blockade resistant EGFR and Kras mouse lung tumors (a) Total lung weight of untreated
1 Supplementary Figure 1. Immune profiles of untreated and PD-1 blockade resistant EGFR and Kras mouse lung tumors (a) Total lung weight of untreated (U) EGFR TL mice (n=7), Kras mice (n=7), PD-1 blockade
Immunobiology Handbook
Immunobiology Handbook Immunobiology Handbook BioLegend is ISO 13485:2003 Certified Toll-Free Tel: (US & Canada): 1.877.BIOLEGEND (246.5343) Tel: 858.768.5800 World-Class Quality Superior Customer Support
The autoimmune disease-associated PTPN22 variant promotes calpain-mediated Lyp/Pep
SUPPLEMENTARY INFORMATION The autoimmune disease-associated PTPN22 variant promotes calpain-mediated Lyp/Pep degradation associated with lymphocyte and dendritic cell hyperresponsiveness Jinyi Zhang, Naima
Supplementary Fig. 1: Ex vivo tetramer enrichment with anti-c-myc beads
Supplementary Fig. 1: Ex vivo tetramer enrichment with anti-c-myc beads Representative example of comparative ex vivo tetramer enrichment performed in three independent experiments with either conventional
MATERIALS. Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL
v Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL Authors S. Kutscher, A. Cosma, MATERIALS REAGENTS: - PBMCs - Culture medium - Costimulating antibodies - Peptide pools including
Simultaneous correlation of cytokine production with Treg and Th17 cell proliferation
Simultaneous correlation of cytokine production with Treg and Th17 cell proliferation Jurg Rohrer, PhD Director, R&D BD Biosciences 23-11773-00 For Research Use Only. Not for use in diagnostic or therapeutic
A step-by-step approach to build and analyze a multicolor panel
Analyze A step-by-step approach to build and analyze a multicolor panel For Research Use Only. Not for use in diagnostic or therapeutic procedures. Alexa Fluor is a registered trademark of Life Technologies
January 25, 2017 Scientific Research Process Name of Journal: ESPS Manuscript NO: Manuscript Type: Title: Authors: Correspondence to
January 25, 2017 Scientific Research Process Name of Journal: World Journal of Gastroenterology ESPS Manuscript NO: 31928 Manuscript Type: ORIGINAL ARTICLE Title: Thiopurine use associated with reduced
Low Avidity CMV + T Cells accumulate in Old Humans
Supplementary Figure Legends Supplementary Figure 1. CD45RA expressing CMVpp65-specific T cell populations accumulate within HLA-A*0201 and HLA-B*0701 individuals Pooled data showing the size of the NLV/HLA-A*0201-specific
Comprehensive evaluation of human immune system reconstitution in NSG. and NSG -SGM3 mouse models toward the development of a novel ONCO-HU
Comprehensive evaluation of human immune system reconstitution in NSG and NSG -SGM3 mouse models toward the development of a novel ONCO-HU xenograft model Aaron Middlebrook, 1 Eileen Snowden, 2 Warren
Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham
Primary Adult Naïve CD4+ CD45RA+ Cells Prepared by: David Randolph (drdrdr@uab.edu) at University of Alabama, Birmingham Goal: To obtain large numbers of highly pure primary CD4+ CD45RO- CD25- cells from
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
T-Cell Research. Novel multicolor flow cytometry tools for the study of CD4 + T-cell differentiation and plasticity
T-Cell Research Novel multicolor flow cytometry tools for the study of CD4 + T-cell differentiation and plasticity A Solid Commitment to Research: Flexible Ways to Study CD4 T-Cell Differentiation and
Cover Page. The handle holds various files of this Leiden University dissertation
Cover Page The handle http://hdl.handle.net/1887/24366 holds various files of this Leiden University dissertation Author: Buddingh, Emilie Pauline Title: Innate immunity in osteosarcoma Issue Date: 2014-03-05
DISCOVER MORE WITH LESS SAMPLE. nanostring.com/3d
nanostring.com/3d DISCOVER MORE WITH LESS SAMPLE. WHY COMPROMISE? ncounter Vantage ASSAYS POWERED BY 3D BIOLOGY TECHNOLOGY Don t let sample volume limit your analytical aspirations. Quantify DNA, RNA,
Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4
Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4 Ankita Garg, Rodney Trout and Stephen A. Spector,,* Department
DURACLONE RE THERE ARE EVENTS YOU CANNOT AFFORD TO MISS
DURACLONE RE THERE ARE EVENTS YOU CANNOT AFFORD TO MISS Your clinical research trial companion For Reseach Use Only - Not for use in Diagnostic procedures THERE ARE EVENTS YOU CANNOT AFFORD TO MISS Rare
Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2*
Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2* 1 Department of Laboratory Medicine - Laboratory of Hematology, Radboud University
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
For surface staining, cells were stained with various markers (supplemental table S1) at room
Supplemental materials and methods. Flow cytometry( details) : For surface staining, cells were stained with various markers (supplemental table S1) at room temperature for 15 minutes, washed with PBS
CD14 + S100A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer
CD14 + S1A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer Po-Hao, Feng M.D., Kang-Yun, Lee, M.D. Ph.D., Ya-Ling Chang, Yao-Fei Chan, Lu- Wei, Kuo,Ting-Yu
sequences of a styx mutant reveals a T to A transversion in the donor splice site of intron 5
sfigure 1 Styx mutant mice recapitulate the phenotype of SHIP -/- mice. (A) Analysis of the genomic sequences of a styx mutant reveals a T to A transversion in the donor splice site of intron 5 (GTAAC
Supplemental Figure 1. IL-3 blockade with Fab CSL362 depletes plasmacytoid dendritic cells (pdcs), but not basophils, at higher doses.
Supplemental Figure 1. IL-3 blockade with Fab CSL362 depletes plasmacytoid dendritic cells (pdcs), but not basophils, at higher doses. Percentage of viable (A) pdcs (Sytox Blue-, Lin1-, HLADR+, BDCA2++)
ABSTRACT INTRODUCTION
/, Vol. 7, No. 34 Inherent low Erk and p38 activity reduce Fas Ligand expression and degranulation in T helper 17 cells leading to activation induced cell death resistance Doureradjou Peroumal 1, Thiruvaimozhi
Commercially available HLA Class II tetramers (Beckman Coulter) conjugated to
Class II tetramer staining Commercially available HLA Class II tetramers (Beckman Coulter) conjugated to PE were combined with dominant HIV epitopes (DRB1*0101-DRFYKTLRAEQASQEV, DRB1*0301- PEKEVLVWKFDSRLAFHH,
Reagents Catalog 4 th Edition Human Mouse Non-Human Primate
Welcome to More Choice Multicolor Antibody Reagents Catalog 4 th Edition Human Mouse Non-Human Primate Choose from our extensive portfolio of high-quality fluorescent-conjugated reagents to build your
BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit (with BD GolgiStop protein transport inhibitor containing monensin) (Cat. No.
Kit Manual BD Cytofix/Cytoperm Fixation/Permeabilization Kit (Cat. No. 554714) BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit (with BD GolgiStop protein transport inhibitor containing monensin)
CELLULAR IMMUNE CORRELATES OF PROTECTION AGAINST
CELLULAR IMMUNE CORRELATES OF PROTECTION AGAINST SYMPTOMATIC PANDEMIC INFLUENZA Saranya Sridhar 1, Shaima Begom 1, Alison Bermingham 2, Katja Hoschler 2, Walt Adamson 3, William Carman 3, Thomas Bean 4,
Supplementary appendix
Supplementary appendix This appendix formed part of the original submission and has been peer reviewed. We post it as supplied by the authors. Supplement to: Kennedy GA, Varelias A, Vuckovic S, et al.
For research or further manufacturing use only. Not for injection or diagnostic procedures.
PRIME-XV T cell Expansion XSFM PRIME-XV T Cell Expansion XSFM is a xeno-free, serum-free medium optimized for the activation and expansion of human T lymphocytes. This medium contains gentamicin and requires
Immunohistochemistry Research Products
Immunohistochemistry Research Products Multiple Tissue Arrays (MTA) Primary and Secondary Antibodies Detection Systems Antigen Retrieval Solutions Tissue Processing Fixatives BD Biosciences Clontech Discovery
Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and
Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and Thy1 in NH cells derived from the lungs of naïve mice.
JAK3 inhibitor administration in vivo in chronically SIV Infected Rhesus Macaques
JAK3 inhibitor administration in vivo in chronically SIV Infected Rhesus Macaques preferentially depletes NK Cells Ladawan Kowawisatsut 1 Kovit Pattanapanyasat 1 Aftab A. Ansari 2 1 Department of Immunology
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.
HIV-infected subjects were recruited from an ongoing clinic-based cohort (SCOPE)
SUPPLEMENTAL METHODS Populations studied HIV-infected subjects were recruited from an ongoing clinic-based cohort (SCOPE) based at the University of California, San Francisco. From this cohort, we recruited
Technical Resources. BD Immunocytometry Systems. FastImmune Intracellular Cytokine Staining Procedures
FastImmune Intracellular Cytokine Staining Procedures BD has developed protocols for the detection of intracellular cytokines in activated lymphocytes and in activated monocytes. The procedures have been
Supplemental Information. Human CD1c + Dendritic Cells Drive. the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF-
Immunity, Volume 38 Supplemental Information Human CD1c + Dendritic Cells Drive the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF- Chun I. Yu Christian Becker Yuanyuan
L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity
Cell Reports Supplemental Information L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity Rebar N. Mohammed, H. Angharad Watson, Miriam Vigar,
Supplementary Table 1: Summary of Reactogenicity Events by Group and Time of Onset in Entebbe
Supplementary Table 1: Summary of Reactogenicity Events by Group and Time of Onset in Group A (n=39) B (n=11) A (n=39) B (n=11) Onset Post-Vaccination Event 3 Minutes 3-14 days Pain 5 (13%) 1 (1%) 6 (15%)
T cell protein tyrosine phosphatase attenuates T cell signaling to maintain tolerance in mice
SUPPLEMENTAL METHODS T cell protein tyrosine phosphatase attenuates T cell signaling to maintain tolerance in mice Florian Wiede 1, Benjamin J. Shields 1, Sock Hui Chew 1, Konstantinos Kyparissoudis 2,
Supplementary Figure Legends. group) and analyzed for Siglec-G expression utilizing a monoclonal antibody to Siglec-G (clone SH2.1).
Supplementary Figure Legends Supplemental Figure : Naïve T cells express Siglec-G. Splenocytes were isolated from WT B or Siglec-G -/- animals that have not been transplanted (n= per group) and analyzed
SUPPLEMENTARY FIGURES
SUPPLEMENTARY FIGURES Supplementary Figure 1: Chemokine receptor expression profiles of CCR6 + and CCR6 - CD4 + IL-17A +/ex and Treg cells. Quantitative PCR analysis of chemokine receptor transcript abundance
Table S1. Viral load and CD4 count of HIV-infected patient population
Table S1. Viral load and CD4 count of HIV-infected patient population Subject ID Viral load (No. of copies per ml of plasma) CD4 count (No. of cells/µl of blood) 28 7, 14 29 7, 23 21 361,99 94 217 7, 11
Supplementary Notes. 2. Nine cell surface markers, CD33, CD20, CD3, CD4, CD7, CD123, CD14, IgM, and HLA-DR,
Supplementary Notes Supplementary Note 1: Mass-tag cellular multiplexing workflow First, the cellular state is preserved by formaldehyde cross-linking. Second, the cells are permeabilized with methanol,
IMMUNOLOGY. Source, Isolate, Culture, And Analyze Immune Cells. Scientists Helping Scientists
IMMUNOLOGY Source, Isolate, Culture, And Analyze Immune Cells Scientists Helping Scientists WWW.STEMCELL.COM TABLE OF CONTENTS Tools For Your Immunology Research 4 Primary Cells: It All Starts with The
Product Datasheet. CD161/NK1.1 Antibody (PK136) NB Unit Size: 0.5 mg. Store at 4C. Do not freeze. Publications: 2
Product Datasheet CD161/NK1.1 Antibody (PK136) NB100-77528 Unit Size: 0.5 mg Store at 4C. Do not freeze. Publications: 2 Protocols, Publications, Related Products, Reviews, Research Tools and Images at:
Roles of calpain-calpastatin system (CCS) in human T cell activation
/, Vol. 7, No. 47 Roles of calpain-calpastatin system (CCS) in human T cell activation Anna Mikosik 1, Aleksandra Jasiulewicz 1, Agnieszka Daca 2, Izabella Henc 2, Joanna E. Frąckowiak 1, Katarzyna Ruckemann-Dziurdzińska
Supplementary Data 1. Alanine substitutions and position variants of APNCYGNIPL. Applied in
Supplementary Data 1. Alanine substitutions and position variants of APNCYGNIPL. Applied in Supplementary Fig. 2 Substitution Sequence Position variant Sequence original APNCYGNIPL original APNCYGNIPL
Use of standardized lyophilized reagents to develop a functional T- cell signature. John F. Dunne, PhD Assoc. Scientific Director BD Biosciences
Use of standardized lyophilized reagents to develop a functional T- cell signature John F. Dunne, PhD Assoc. Scientific Director BD Biosciences Functional T cell responses to tumor antigens in breast cancer
CHARACTERIZING CHRONIC LYMPHOCYTIC LEUKEMIA FOR NOVEL BIOMARKERS OF CD40 RESPONSIVENESS USING INTRACELLULAR FLOW CYTOMETRY CHRISTOPHER M.
CHARACTERIZING CHRONIC LYMPHOCYTIC LEUKEMIA FOR NOVEL BIOMARKERS OF CD40 RESPONSIVENESS USING INTRACELLULAR FLOW CYTOMETRY by CHRISTOPHER M. BOODY A thesis submitted to the Graduate School-New Brunswick
Nature Immunology: doi: /ni Supplementary Figure 1. Huwe1 has high expression in HSCs and is necessary for quiescence.
Supplementary Figure 1 Huwe1 has high expression in HSCs and is necessary for quiescence. (a) Heat map visualizing expression of genes with a known function in ubiquitin-mediated proteolysis (KEGG: Ubiquitin
Supplementary Figure 1. Gating strategy for leukemic blasts and normal developing B
Supplementary Information Supplementary Figure 1. Gating strategy for leukemic blasts and normal developing B cells. (a) Gating strategy for lineage-negative blasts (B + Lin - cells), the starting population
Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or
Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or control nontargeting sirnas. At 90 hr after transfection,
In Vivo Administration of a JAK3 Inhibitor during Acute SIV Infection Leads to Significant Increases in Viral Load during Chronic Infection
In Vivo Administration of a JAK3 Inhibitor during Acute SIV Infection Leads to Significant Increases in Viral Load during Chronic Infection Yoshiaki Takahashi, Emory University Siddappa N. Byrareddy, Emory
Supplementary Figure S1. DD2 reactivity on human tonsils and analysis of slandc proliferation. Sections are from a representative human tonsil (a-f;
Supplementary Figure S1. DD2 reactivity on human tonsils and analysis of slandc proliferation. Sections are from a representative human tonsil (a-f; n=10) and a representative M- TDLN (g-h; n=5) and stained
MBL in Blood Donors: An Extended Immunophenotypic Analysis of CLL-like, CD5+ MBL.
1 th Canadian CLL Meeting Winnipeg, Manitoba Inn at the Forks September 18-19, 214 MBL in Blood Donors: An Extended Immunophenotypic Analysis of CLL-like, CD5+ MBL. Fatima Abbasi 1, Justin Meskas 2, Ryan
Functional and Phenotypic Analysis of B- and T- Helper Cells in Rhesus Macaques and African Green Monkeys
University of Wisconsin Milwaukee UWM Digital Commons Theses and Dissertations December 2015 Functional and Phenotypic Analysis of B- and T- Helper Cells in Rhesus Macaques and African Green Monkeys Sahar
A Multifaceted Immunomonitoring to Identify Predictive Biomarkers for the Clinical Outcome of Immunotherapy-Treated Melanoma Patients
A Multifaceted Immunomonitoring to Identify Predictive Biomarkers for the Clinical Outcome of Immunotherapy-Treated Melanoma Patients Immunotherapy Biomarkers: Overcoming the Barriers NIH, Bethesda, April
Supplementary Figure 1. Enhanced detection of CTLA-4 on the surface of HIV-specific
SUPPLEMENTARY FIGURE LEGEND Supplementary Figure 1. Enhanced detection of CTLA-4 on the surface of HIV-specific CD4 + T cells correlates with intracellular CTLA-4 levels. (a) Comparative CTLA-4 levels
CONTRACTING ORGANIZATION: Johns Hopkins University School of Medicine Baltimore, MD 21205
AD Award Number: DAMD7---7 TITLE: Development of Artificial Antigen Presenting Cells for Prostate Cancer Immunotherapy PRINCIPAL INVESTIGATOR: Jonathan P. Schneck, M.D., Ph.D. Mathias Oelke, Ph.D. CONTRACTING
Interleukin-2-Dependent Allergen-Specific Tissue-Resident Memory Cells Drive Asthma
Immunity Supplemental Information Interleukin-2-Dependent Allergen-Specific Tissue-Resident Memory Cells Drive Asthma Brian D. Hondowicz, Dowon An, Jason M. Schenkel, Karen S. Kim, Holly R. Steach, Akshay
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
IFN- -rich environment programs dendritic cells toward silencing of cytotoxic immune responses
Article IFN- -rich environment programs dendritic cells toward silencing of cytotoxic immune responses Urban Švajger,*,,1 Nataša Obermajer,, and Matjaž Jeras, *Blood Transfusion Centre of Slovenia, Ljubljana,
Immunobiology Handbook
Immunobiology Handbook Immunobiology Handbook BioLegend is ISO 9001:2008 and ISO 13485:2003 Certified Toll-Free Tel: (US & Canada): 1.877.BIOLEGEND (246.5343) Tel: 858.768.5800 biolegend.com World-Class
Cytotoxicity assays. Rory D. de Vries, PhD 1. Viroscience lab, Erasmus MC, Rotterdam, the Netherlands
Cytotoxicity assays Rory D. de Vries, PhD 1 1 Viroscience lab, Erasmus MC, Rotterdam, the Netherlands Anti-influenza immunity Humoral / CD4+ / CD8+ / NK? Function of CTL Elimination of virus-infected cells?
x Lymphocyte count /µl CD8+ count/µl 800 Calculated
% Lymphocyte in CBC A. 50 40 30 20 10 Lymphocyte count /µl B. x10 3 2.5 1.5 C. 50 D. 1000 % CD3+CD8+ Cells 40 30 20 Calculated CD8+ count/µl 800 600 400 200 10 0 #61 #63 #64 #65 #68 #71 #72 #75 Figure
CyTOF analyses in rheumatoid arthritis. Deepak Rao, MD PhD Rheumatology, Immunology, Allergy, BWH
CyTOF analyses in rheumatoid arthritis Deepak Rao, MD PhD Rheumatology, Immunology, Allergy, BWH Agenda: CyTOF analyses Analysis of synovial tissue Panel for analysis of synovial cells Comparison of CyTOF
Supplementary Figure 1. Example of gating strategy
Supplementary Figure 1. Example of gating strategy Legend Supplementary Figure 1: First, gating is performed to include only single cells (singlets) (A) and CD3+ cells (B). After gating on the lymphocyte
PBMC were separated using Ficoll-Hypaque (Sigma-Aldrich) and stained with anti -CD19 PE and -
Supplemental Methods CLL cell preparation and culture PBMC were separated using Ficoll-Hypaque (Sigma-Aldrich) and stained with anti -CD19 PE and - CD5 FITC antibodies (Miltenyi Biotec). When the CD19
Chapter 5. Enhanced differentiation of B-cells towards immunoglobulin-secreting cells in rheumatoid arthritis
Enhanced differentiation of B-cells towards immunoglobulin-secreting cells in rheumatoid arthritis Teng YKO Levarht EWN Huizinga TWJ Toes REM van Laar JM Submitted Abstract Objectives To investigate the
This information is current as of November 17, 2018.
This information is current as of November 17, 2018. Supplementary Material References Subscription Permissions Email Alerts T Cell-Signaling Network Analysis Reveals Distinct Differences between CD28