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High Throughput Cytotoxicity

Cytolytic activity is an important process for eliminating intracellular pathogens and cancer cells.  This process is accomplished through various immune effector mechanisms including Natural Killer (NK) leukocytes. Natural killer activity is an innate immune response accomplished by non-specifically lysing infected cells 


 
KEY BENEFITS:
• No radioactive compounds so potential health risk is eliminated.
• Reduction in spontaneous leakage, which has resulted in high background readings in the past.
• Cytolytic activity is measured at a single cell level by flow cytometry

EACH KIT INCLUDES:
Kit sizes of Cy150, Cy250, Cy500 or Cy1000 tests including materials for optimization of effector/target cell ratios (see line graph above).
• CFSE fluorescent membrane stain for labeling the target cells
• 7-AAD for Live-Dead Stain 

Cytotoxicity Manual

       References:  

1. Lee-MacAry, A.E., Ross, E.L, Davies, D., and Wilkinson, R.W., 2001. Development of a novel flow cytometric cell-mediated cytotoxcity assay using the fluorophores PKH-26 and TO-PRO-3 iodide. J. Immunol. Methods 252: 83-92.

2. Radosevic, K., Garritsen, H.S.P., Van Graft, M., De Grooth, B.G. and Greve, J., 1990. A simple and sensitive flow cytometric assay for the determination of the cytotoxic activity of human natural killer cells. J. Immunol. Methods 135:81

3. Hatam, L,. Schuval, S., Bonagura, V.R., 1994. Flow cytometric analysis of natural killer cell function as a clinical assay. Cytometry 16:59.

4. Lowdell, M.W., Ray, N., Graston, R., Corbett, T., Deane, M. and Prentice, H.G., 1997. The in vitro detection of anti-leukemia-specific cytotoxicity after autologous bone marrow transplantation for acute leukemia. Bone Marrow Transplant. 19:891.

5. Van Hooijonk, C.A.E.M., Glade, C.P. and Van Erp, P.E.J., 1994. TO-PRO-3 Iodide: a novel HeNe laser-excitable DNA stain as an alternative for propidium iodide in multiparameter flow cytometry. Cytometry 17:185.

6. O’Brien, M.C. and Bolton, W.E., 1995. Comparison of cell viability probes compatible with fixation and permeabilization for combined surface and intracellular  staining in flow cytometry. Cytometry 19:243.   

 7. Olin, M., K. Choi, J. Lee, and T. W. Molitor. 2004. Gamma Delta T-lymphocytes cytolytic activity against Mycobacterium tuberculosis analyzed by flow cytometry. J Immunol Methods, 297:1-11..

 


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AQ Molecular and Immunochemical