7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit

Cayman Chemical Item Number 600120

7-Amino Actinomycin D/5-(6)-Carboxyfluorescein Diacetate Succinimidyl Ester

Description

Cell-mediated cytotoxicity is characterized by cytolysis of a target cell by effector cells such as cytotoxic T lymphocytes or natural killer cells. It is a major part of the immune system in which the body protects itself from attack by invading pathogens and is fundamental in maintaining homeostasis of the immune system. Dysfunction of cytotoxic T lymphocytes or natural killer cells results in pathological conditions such as multiple sclerosis and cancer.¹ Cell-mediated cytotoxicity is a complex, multistep process which is mediated by several different pathways, including the granule exocytosis pathway and the Fas pathway. The interplay between these pathways provides opportunities for therapeutic interventions to control autoimmune diseases and graft versus host disease.² Cell-mediated cytotoxicity has typically been assessed using the ⁵¹chromium (⁵¹Cr) release assay, in which the target cells are labeled with ⁵¹Cr. This method has disadvantages including the use of radioisotopes and low sensitivity. More sensitive and convenient methods have been developed using green fluorescent probes such as 5-(6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) to label target cells, in combination with red fluorescent probes such as 7-AAD or propidium iodide to label dead effector and dead target cells. CFSE passively diffuses into cells and its acetate groups are cleaved by intracellular esterases to yield higly fluorescent carboxyfluorescein succinimidyl ester. 7-AAD and propidium iodide are fluorescent dyes which are excluded from live cells but penetrate dead or damaged cells to label DNA. The labeling distinguishes four populations of cells: 1) living target cells in green, 2) dead target cells in green and red, 3) dead effector cells in red, and 4) live effector cells, which remain unstained. Thus, the cytotoxicity of the effector cells can be evaluated quantitatively. Cayman’s 7-AAD/CFSE Cell-Mediated Cytotoxicity Assay Kit employs CFSE to label target cells and 7-AAD to label dead cells. This kit provides a convenient tool to quantify the cytotoxic effect of immune effector cells at a single cell level. The kit provides sufficient reagents to effectively treat/stain 10 plates worth of cells irrespective of the number of wells per plate.

1 Benczur, M., Petrányi, G.G., Pálffy, G., et al. Dysfunction of natural killer cells in multiple sclerosis: A possible pathogenetic factor. Clin Exp Immunol 39 657-662 (1980).

2 Russell, J.H., and Ley, T.J. Lymphocyte-mediated cytotoxicity. Annu Rev Immunol 20 323-370 (2002).

Synonyms

7-Amino Actinomycin D/5-(6)-Carboxyfluorescein Diacetate Succinimidyl Ester

Stability

1 year

Storage

-20°C

Shipping

Wet ice in continental US; may vary elsewhere

Background Reading

Benczur, M., Petrányi, G.G., Pálffy, G., et al. Dysfunction of natural killer cells in multiple sclerosis: A possible pathogenetic factor. Clin Exp Immunol 39 657-662 (1980).

Russell, J.H., and Ley, T.J. Lymphocyte-mediated cytotoxicity. Annu Rev Immunol 20 323-370 (2002).

600120-1ea

Cell-Based Assay Buffer Tablet

4 ea

Cell-Based Assay 7-AAD Staining Stock Solution (1,000X)

2 × 1 ea

CFSE Stock Solution

1 ea

Size

Price

Quantity

Subtotal