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Apoptotic, Necrotic, and Healthy Cells Quantification Kit

The kit allows simultaneous quantification of apoptotic (green), necrotic (red) and all cells (blue) within the same cell population by flow cytometry or fluorescence microscopy.

30018 Jurkat, staurosporine
Catalog #

30018

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50 assays

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Product Description

Apoptosis and necrosis are two major processes by which cells die. Apoptosis is an active, genetically regulated disassembly of the cell from within. During apoptosis, phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. The human anticoagulant Annexin V is a 35 kDa, Ca2 -dependent phospholipid-binding protein with a high affinity for PS. Annexin V labeled with fluorescein (FITC) (λExEm = 492/514 nm) can identify apoptotic cells in green by binding to PS exposed on the outer leaflet. Necrosis normally results from a severe cellular insult, resulting in loss of plasma membrane integrity. Ethidium homodimer III (EthD-III) is a highly positively charged nucleic acid probe, which is impermeant to live cells or apoptotic cells, but stains necrotic cells with red fluorescence (λExEm (with DNA) = 528/617 nm). EthD-III is a superior alternative to propidium iodide (PI) or ethidium homodimer I because of its significantly higher affinity for DNA and higher fluorescence quantum yield. The kit also includes the membrane-permeant blue fluorescent DNA dye Hoechst 33342 (λExEm (with DNA) = 350/461nm) for visualizing all cells. The kit allows simultaneous quantification of apoptotic (green), necrotic (red) and all cells (blue) within the same cell population by flow cytometry or fluorescence microscopy.

Kit Components:

  • FITC-Annexin V
  • Ethidium Homodimer III
  • Hoechst 33342
  • 5X binding Buffer

Features:

  • Simple: Simultaneous detection of apoptotic, necrotic and healthy cells in a single assay
  • Fast: A single 15-30 min staining step
  • Sensitive: EthD-III is a superior alternative to both PI and EthD-I for staining necrotic cells.

Also see the Apoptosis and Necrosis Quantitation Kit Plus (catalog no. 30065) and Apoptotic, Necrotic, and Healthy Cells Quantitation Kit Plus (catalog no. 30066), which feature Annexin V conjugated to Biotium’s superior green fluorescent dye CF™488A, which is brighter and more photostable than FITC.

References

1. Physical Biology (2011) doi:10.1088/1478-3975/8/1/015006

2. Pharmacological Reports 63, 139 (2011).

3. International Journal of Molecular Sciences 9, 56 (2008)

.

4. Science 300, 91 (2003).

5. J Neurosci Methods. 36, 229 (1991).

6. FEBS Lett 203, 99 (1986).

7. Biosci. Rep. 7, 289 (1987).

 

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Support & Faq

Apoptosis, Necrosis, and Cell Viability Kits

MTT and XTT are colorimetric based assays, while resazurin can be measured using colorimetric or fluorescence detection. MTT is not a soluble product, so the cells must be lysed to solubilize the formazan salt before absorbance can be measured. XTT and resazurin do not require cell lysis, allowing kinetic monitoring of the same samples at different timepoints.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

Check to make sure your cell densities are in the linear range of the cell viability assay. Too few cells may fall below the limit of detection, and too many cells may saturate the OD. Perform an initial experiment to check the best linear range for your cell type and experimental set-up. You may need to vary the cell density assay incubation time. The kit protocols provide general guidelines and may need to be optimized for your experimental system.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

Resazurin is reduced by cells to the fluorescent product resorufin. Resorufin can be reduced further to a non-fluorescent compound. Therefore the densest wells may have lowest fluorescence due to over-reduction of the substrate. Please see the product information sheet for more details. The kit protocol provides general guidelines and may need to be optimized empirically for your experimental system. You may need to vary cell density or assay incubation time to ensure that your samples fall in the linear range of the kit.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

← FAQs

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Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions. Expedited shipment may be requested at time of checkout. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.