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Apoptosis and Necrosis Quantitation Kit Plus

The Apoptosis & Necrosis Quantitation Kit Plus provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy

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Apoptosis/viability marker

Phosphatidylserine/Annexin V, Dead cell stain, Apoptosis/necrosis assay

For live or fixed cells

For live/intact cells

Detection method/readout

Fluorescence microscopy, Flow cytometry

Assay type/options

Endpoint assay

Colors

Green/Red

Size
Catalog #
price
Qty
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 leaflet of the plasma membrane for phagocytic cell recognition. The human anticoagulant Annexin V is a 35 kDa, Ca2 -dependent phospholipid-binding protein with high affinity for PS. Annexin V labeled with CF®488A (Abs/Em: 490/515 nm) stains apoptotic cells green by binding to PS exposed on the cell surface. CF®488A is spectrally similar to fluorescein (FITC), with much brighter and more photostable fluorescence.

Necrosis normally results from severe cellular insult. Both internal organelle and plasma membrane integrity are lost, resulting in the release of cellular contents into the surrounding environment. Ethidium homodimer III (EthD-III) is a highly positively charged nucleic acid probe, which is impermeant to live cells, but stains cells with compromised membrane integrity with red fluorescence (Abs/Em (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 Apoptosis & Necrosis Quantitation Kit Plus provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy.

Kit Components:

  • CF®488A-Annexin V
  • Ethidium homodimer III
  • 5X binding buffer

Features:

  • Simple: Simultaneous detection of both apoptotic and necrotic cells in a single assay
  • Fast: A single 15-30 min staining step
  • Sensitive: CF®488A-Annexin V is brighter and more photostable than FITC-Annexin V, while EthD-III is a superior alternative to both PI and EthD-I for staining necrotic cells.

References

1. Cvjeticanin, T., et al. T Cells Cooperate with Palmitic Acid in Induction of Beta Cell Apoptosis. BMC Immunology doi:10.1186/1471-2172-10-29 (2010).

2. Biol. of Reproduction (2016), doi:10.1095/biolreprod.115.134924

 

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Support & Faq

Apoptosis, Necrosis, and Cell Viability Kits

Fixation is not recommended after staining with the Viability/Cytotoxicity Assay Kit (cat. # 30027) as the dead cell dye, EthD-III will transfer to all cells and not maintain dead cell-specific staining.

Viability qPCR (vPCR) can be an alternative strategy. Using dead cell-specific dyes, PMA (cat. # 40019) or PMAxx (cat. # 40069) that covalently modify DNA (of only dead cells) after photo-crosslinking, a simple quantitative PCR (qPCR) amplification is used to selectively amplify live-cell DNA. Learn more about the vPCR technology.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

The ViaFluor® SE dyes are susceptible to hydrolysis. Ideally the DMSO stock solution should be prepared on the day of use, with the intent that each vial supplied is for single use. If smaller aliquots are desired, it would be best to aliquot the initial DMSO stock, at the time of reconstitution, into single use quantities and store at -20C, protected from light and moisture (desiccated), for up to a month. However, activity may be reduced over time. The dyes should only be added to aqueous buffer immediately before staining. The aqueous solution cannot be stored for reuse.

Category: Apoptosis, Necrosis, and Cell Viability Kits

← FAQs

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

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Shipping & Handling Disclaimer: Shipping and handling methods are calculated based on item temperature storage recommendation. Large, multi-item orders may be assessed additional fees. Shipping to Puerto Rico may be assessed additional fees. A Biotium customer service representative will contact you if additional fees are required. For expedited shipping, please request in the Order Notes section of the checkout page. 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.