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Product Details
| Cat # +Size | K315-100 |
|---|---|
| Size | 100 assays |
| 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 |
| Features & Benefits | • Direct method to measure cell-mediated cytotoxicity. • Does not require cell lysis. • Provides multi-parametric analysis. |
| Kit Components | • Cytotoxicity Assay Buffer • 7-AAD Staining Solution • CFSE Staining Solution |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
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. BioVision’s 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. The flow cytometry-based method provides robust data and enables multi-parametric analysis.
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How to analyse the data?
Any live cell is quantified by CFSE stain and apoptotic/necrotic cells by 7AAD stain. In the dual format, you can analyse all populations. Untreated live cells: high on CFSE axis, low on 7AAD axis – right lower quadrantTreated live but apoptotic cells: high on both axes on the right higher quadrant
Does it require BSA?
Cells are grown in media containing FBS
Can this kit be used with an imaging system instead of flow cytometry? What excitation/emission wavelengths are used for detection?
CFSE has a wavelength excitation maximum of 492 nm and emission maximum of 517 nm. 7 AAD has en excitation max of 488 nm and emission max is 665 nm. These filters can be used to image healthy versus Apoptotic cells. See reference here: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3466660/
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
