Cell-Mediated Cytotoxicity Assay Kit (Fluorometric)

• Reactivity: Hu, Mu, Rt, Ma
• Applications: Flow
• Suitable Sample Type: Adherent and suspension cells

Cell-Mediated Cytotoxicity Assay Kit (Fluorometric) Summary

• Description: Direct method to measure cell-mediated cytotoxicity. Does not require cell lysis and provides multi-parametric analysis.
  
  • Detection method: Fluorescence (FL1 channel and FL3 channel)
  • Applications: Measurement of cell cytotoxicity in response to drug or toxin treatment. Quantification of the cytotoxic effect of immune effector cells on target cells. Assessment of physiological mediators and antibodies that affect cell cytotoxicity.
• Kit Type: Assay Kit (Fluorometric)

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
• Application Notes: Staining treated cells with CFSE alone and 7-AAD alone to choose the proper instrument gating set up. Keep unstained control cells (i.e. without CFSE or 7-AAD staining) suspended in Cytotoxicity Assay Buffer for both treated and untreated samples to set up the flow cytometer. CFSE cell staining is measured in the FL1 channel and 7-AAD cell staining is measured in the FL3 channel. To ensure that only proper target cells are gated, use a side scatter versus FL-1 plot.

Packaging, Storage & Formulations

• Storage: Store at -20C.

Background

Cell-mediated cytotoxicity is an important phenomenon characterized by cytolysis of a compromised cell in the body by immune system. Activation of immune system leads to removal of target cells infected by pathogens or transformed cells/cancer cells. This process is mediated by antibody-dependent cell-mediated cytotoxicity (ADCC), complement-mediated cytotoxicity, or lymphocyte-mediated cytotoxicity. This Cell-Mediated Cytotoxicity Assay Kit contains carboxyfluorosuccinimide ester (CFSE), a green fluorescent probe that labels live target cells and 7-aminoactinomycin D (7-AAD), a red fluorescent probe that labels late apoptotic and necrotic target cells killed in the cytotoxicity assay. This assay does not require cell lysis and provides a direct measurement of cytotoxicity instead of indirect indicators such as release of ATP or lactate dehydrogenase activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.