Product Datasheet. Annexin V Apoptosis Kit [FITC] NBP Tests. Unit Size: 100 Tests. Store at 4C. Do not freeze.

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1 Product Datasheet Annexin V Apoptosis Kit [FITC] NBP Tests Unit Size: 100 Tests Store at 4C. Do not freeze. Publications: 20 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 8/21/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NBP Tests Annexin V Apoptosis Kit [FITC] Product Information Unit Size Concentration Storage Conjugate 100 Tests Concentration is not relevant for this product. Please see the protocols for proper use of this product. Store at 4C. Do not freeze. FITC Page 1 of 5 v.20.1 Updated 8/21/2018 Product Description Description Gene ID 308 Gene Symbol Species Reactivity Notes Kit Components Notes Annexin V Apoptosis Kit [FITC] can identify apoptosis at an earlier stage than kits based on DNA fragmentation in the nucleus. However, it is like most assays and has limitations. Since Annexin V staining precedes the loss of membrane integrity which accompanies the later stages of cell death resulting from either apoptotic or necrotic processes, staining with Annexin V-FITC is typically used in conjunction with a live/dead dye such as propidium iodide (PI) or 7-Amino- Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells (PI negative, Annexin V-FITC positive) from dead cells (PI positive, AnnexinV-FITC positive). Viable cells with intact membranes exclude PI, whereas the membranes of dead and damaged cells are permeable to PI. For this reason, Annexin V staining has to be performed on live cells as opposed to assays which require para- formaldehyde/ethanol fixed cells. The Annexin V assay works in the following manner: Cells that are viable are both Annexin V-FITC and PI negative. While cells that are in early apoptosis are Annexin V-FITC positive and PI negative and cells that are in late apoptosis or already dead are both FITC Annexin V and PI positive. This assay does not distinguish between cells that have undergone apoptotic death versus those that have died as a result of a necrotic pathway because the dead cells will stain with both Annexin V-FITC and PI (see image). However, when apoptosis is measured over time, cells can be often tracked from Annexin V- FITC and PI negative (viable, or no measurable apoptosis), to Annexin V-FITC positive and PI negative (early apoptosis, membrane integrity is present) and finally to Annexin V- FITC and PI positive (end stage apoptosis and death). The movement of cells through these three stages suggests apoptosis. In contrast, a single observation indicating that cells are both Annexin V-FITC and PI positive, in of itself, reveals less information about the process by which the cells underwent their demise. For this reason, it is a good idea to analyze samples from multiple time points. ANXA5 Human, Mouse, Rat Human reactivity reported in scientific literature (PMID: ) Mouse reactivity reported in scientific literature (PMID: ). Rat reactivity reported in scientific literature (PMID: ) Annexin V-FITC, 10X Binding Buffer, Propidium Iodide (PI) Solution, 10X PBS Solution, Compensation Control Cells ADDITIONAL ITEMS REQUIRED (NOT INCLUDED IN THE KIT) Distilled H2O Flow Cytometer Experimental Cells Centrifuge Positive Control Cell Line (recommended, not required) Product Application Details

3 Applications Recommended Dilutions Application Notes Page 2 of 5 v.20.1 Updated 8/21/2018 Flow Cytometry, Flow (Cell Surface), Immunocytochemistry/Immunofluorescence Flow Cytometry, Immunocytochemistry/Immunofluorescence, Flow (Cell Surface) Use in Flow cell surface reported in scientific literature (PMID ) Use the Control Cells provided to set up compensation and quadrants. The Control Cells are positive for both Annexin V-FITC and PI. The basal level of apoptosis and necrosis varies considerably within a give cell population. Even in the absence of induced apoptosis, most cell populations will contain a minor percentage of cells that are positive for apoptosis ((Annexin V- FITC positive, PI negative) and dead, necrotic, or in the late stages of apoptosis (Annexin V-FITC positive, PI positive). Thus an untreated cell population is used to define the basal level of apoptotic and dead cells. Determine the percentage of cells that have been induced to undergo apoptosis by subtracting the percentage of apoptotic cells in the untreated from the treated population. Since cell death is the eventual outcome of cells undergoing apoptosis, cells in the late stages of apoptosis will have a damaged membrane and stain positive for PI as well as for Annexin V-FITC. Consequently cells which have undergone necrosis are not distinguishable from those which have undergone apoptosis. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID ). Use in FLOW cytometry reported in scientific literature ( PMID ) Images Flow Cytometry: Annexin V Apoptosis Kit [FITC] [NBP ] - Controls run before the experimental samples are analyzed, to adjust the fluorescent compensation on the flow cytometer due to the FITC signal from the Annexin V-FITC bleeding into the Propidium Iodide signal. Flow Cytometry: Annexin V Apoptosis Kit [FITC] [NBP ] - Adherent RAW cells treated with actinomycin-d, detached with a cell detachment solution and stained with the Annexin V Apoptosis Kit [FITC]. Publications Basset C, Bonnet-Magnaval F, Navarro MG, Touriol C et al. Api5 a new cofactor of estrogen receptor alpha involved in breast cancer outcome Oncotarget 2017 Aug 08 [PMID: ] (Flow, Human) Hartung F, Pardo LA. Guiding TRAIL to cancer cells through Kv10.1 potassium channel overcomes resistance to doxorubicin Eur. Biophys. J Jun 27 [PMID: ] (ICC/IF, Human)

4 Esposito CL, Nuzzo S, Kumar SA et al. A combined microrna-based targeted therapeutic approach to eradicate glioblastoma stem-like cells J Control Release 2016 Jul 21 [PMID: ] (FLOW, Human) Chen Y, Huang H, Yao C et al. Antitumor activity of combined endostatin and thymidine kinase gene therapy in C6 glioma models Cancer Med 2016 Jul 01 [PMID: ] Huang CL, Chao CC, Lee YC et al. Paraquat Induces Cell Death Through Impairing Mitochondrial Membrane Permeability. Mol. Neurobiol May 07 [PMID: ] (ICC/IF, FLOW, Rat) Annexin V-FITC-conjugated apoptosis kit was used for analyzing apoptotic cells in experiments involving PC12 cells. The cells with or without 24 hours of Paraquat/PQ treatments (+ dibucaine or cyclosporin A treatments) were subjected to Annexin V-FITC-staining followed by microscopgy imaging or flow cytometric analysis (Fig 1D). Xue Y, Wu L, Liu Y et al. ENTPD5 Induces Apoptosis in Lung Cancer Cells via Regulating Caspase 3 Expression PLoS ONE 2015 Mar 21 [PMID: ] (FLOW, Human) Song W, Soo Lee S, Savini M et al. Ceria nanoparticles stabilized by organic surface coatings activate the lysosomeautophagy system and enhance autophagic clearance. ACS Nano Oct 28 [PMID: ] (FLOW, Human) Annexin V - FITC Apoptosis Kit (Imgenex's CytoGLO Annexin V-FITC Apoptosis Detection Kit 10085K) used for detecting - Annexin V binding affinity change (%) in HeLa/TFEB cells treated or not with CeO2 nanoparticles or taxol for 24 h (Figure 3A). Wang F, Segatori L. Remodeling the proteostasis network to rescue glucocerebrosidase variants by inhibiting ERassociated degradation and enhancing ER folding. PLoS One [PMID: ] (FLOW, Human) Annexin V - FITC Apoptosis Kit (Imgenex's CytoGLO Annexin V-FITC Apoptosis Detection Kit ) used for detecting - Annexin V binding in lacidipine and EerI treated GD cells, Gaucher?s disease /GD patient-derived fibroblasts homozygous for the L444P (1448T>C) mutation (GM10915). FLOW data is shown in Figure 3A. Wang F, Song W, Brancati G, Segatori L. Inhibition of endoplasmic reticulum-associated degradation rescues native folding in loss of function protein misfolding diseases. J Biol Chem [PMID: ] (FLOW, Human) Annexin V - FITC Apoptosis Kit (Imgenex's CytoGLO Annexin V-FITC Apoptosis Detection Kit ) used for detecting apoptosis induction in L444P GC patient-derived fibroblasts treated or not with ER-associated degradation / ERAD inhibitors (FIGURE 8) Wang F, Chou A, Segatori L. Lacidipine remodels protein folding and Ca 2+ homeostasis in Gaucher's disease fibroblasts: a mechanism to rescue mutant glucocerebrosidase. Chem Biol [PMID: ] (FLOW, Human) Annexin V - FITC Apoptosis Kit (Imgenex's CytoGLO Annexin V-FITC Apoptosis Detection Kit ) used for detecting Annexin V binding in experiments involving L444P Gaucher's disease patient-derived fibroblasts. Skivka LM, Fedorchuk OG, Bezdeneznykh NO. The effect of antineoplastic drug NSC on immunogenicity of B16 melanoma. J Exp Integr Med 2014 Mar 24 (Flow-CS, Mouse) Metastatic mlanoma cells, Fig 2. Narendrula R. rrna Disruption: A Predictive Marker of Response to Taxane Chemotherapy. Thesis 2014 (Flow-CS, Human) A2789 human ovarian carcinoma cell line, Fig 12. More publications at Page 3 of 5 v.20.1 Updated 8/21/2018

5 Procedures MSDS for Non Hazardous Product (NBP ) Material Safety Data Sheet for Non Hazardous Products Page 4 of 5 v.20.1 Updated 8/21/2018 Hazard Information Chemical Name: Non hazardous products. Chemical Formula: N/A CAS Number: N/A EEC-No: N/A Hazard Identification None First Aid Measures Eye Contact: None Skin Contact: None Inhalation: None Ingestion: None Accidental Release Measures This product either does not contain hazardous constituents or the concentration of all chemical constituents are below the regulatory threshold limits described by Occupational Safety Health Administration Hazard Communication Standard 29 CFR and the European Directive 91/155/EEC. 88/379/EEC, and 67/546/EEC. Handling and Storage Exposure Controls / Personal Protection Other Precautions: None Physical and Chemical Properties Form: N/A Color: N/A Odor: N/A Melting Point: N/A Boiling Temperature: N/A Density: N/A Vapor Pressure: N/A Solubility in Water: N/A Flash Point: N/A Explosion limits: N/A Ignition Temperature: N/A

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