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> Basic Cytotoxicity Test BioAssay™ Kit

Basic Cytotoxicity Test BioAssay™ Kit

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Référence 474004-125T

Conditionnement : 125Tests

Marque : US Biological

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474004 Basic Cytotoxicity Test BioAssay™ Kit

Applications
FC
Shipping Temp
Blue Ice
Storage Temp
4°C/-20°C

Sample Type:
Cell culture

Reagent Name:
7-AAD, CFSE

Target:
Cell-mediated cytotoxicity

Excitation/Emission:
7-AAD–546nm/647nm; CFSE–492nm/520nm

Method of Analysis:
Flow cytometry

Intended Use:
Basic Cytotoxicity Test Assay is a single-tube, dual-color assay for determining cytotoxicity by flow cytometry. The BioAssay™ employs a green fluorescent cellular stain, CFSE, to label target cells and the red live/dead viability dye, 7-AAD, to identify the dead cells present in the cytotoxicity BioAssay™ samples. Analyze your results using a flow cytometer.

Test Principle:
Cytolytic activity is an important process for eliminating intracellular pathogens and cancer cells. Basic Cytotoxicity Assay allows you to assess cytolytic activity in cell culture.

The Basic Cytotoxicity BioAssay™ includes two fluorescent reagents: CFSE and 7-AAD. CFSE, a green fluorescing cellular stain, is utilized to identify the target cell population. The unstained effector cells are added and incubated with the target cells. 7-AAD, a red fluorescing live/dead stain, is then added to stain all necrotic cells red by binding to the DNA of membrane-compromised cells. The target cell and effector cell populations can then be easily distinguished, and flow cytometry easily identifies live and necrotic cells.

Kit Components:
125 tests:
CFSE target cell stain, 1 vial
7-AAD vital stain, 1 vial
10X Assay Buffer, 1x30mL

250 tests:
CFSE target cell stain, 1 vial
7-AAD vital stain, 2 vials
10X Assay Buffer, 1x60mL

Storage and Stability:
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Applications
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. Toxicity and Hazards: All products should be handled by qualified personnel only, trained in laboratory procedures.
References
1. Lee-MacAry, A. E. et al. Development of a novel flow cytometric|cell-mediated cytotoxicity assay using the fluorophores PKH-26 and|TO-PRO-3 iodide. J Immunol Methods 252, 83-92 (2001).|2. Slezak, S. E. & Horan, P. K. Cell-mediated cytotoxicity. A highly sensitive|and informative flow cytometric assay. J Immunol Methods 117,|205-214 (1989).|3. Radosevic, K., Garritsen, H. S., Van Graft, M., De Grooth, B. G. & Greve,|J. A simple and sensitive flow cytometric assay for the determination|of the cytotoxic activity of human natural killer cells. J Immunol|Methods 135, 81-89 (1990).|4. Hatam, L., Schuval, S. & Bonagura, V. R. Flow cytometric analysis of|natural killer cell function as a clinical assay. Cytometry 16, 59-68,|doi:10.1002/cyto.990160109 (1994).|5. Lowdell, M. W. et al. The in vitro detection of anti-leukaemia-specific|cytotoxicity after autologous bone marrow transplantation for acute leukaemia. Bone Marrow Transplant 19, 891-897, doi:10.1038/|sj.bmt.1700756 (1997).|6. O’Brien, M. C. & Bolton, W. E. Comparison of cell viability probes|compatible with fixation and permeabilization for combined surface|and intracellular staining in flow cytometry. Cytometry 19, 243-255,|doi:10.1002/cyto.990190308 (1995).|7. Van Hooijdonk, C. A., Glade, C. P. & Van Erp, P. E. TO-PRO-3 iodide: a|novel HeNe laser-excitable DNA stain as an alternative for propidium|iodide in multiparameter flow cytometry. Cytometry 17, 185-189,|doi:10.1002/cyto.990170212 (1994).|8. Olin, M. R., Hwa Choi, K., Lee, J. & Molitor, T. W. Gammadelta Tlymphocyte|cytotoxic activity against Mycobacterium bovis analyzed|by flow cytometry. J Immunol Methods 297, 1-11, doi:10.1016/j.|jim.2004.10.002 (2005).

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