The Apoptotic, Necrotic and Healthy Cells Quantification Kit Plus enables the simultaneous and convenient detection and quantification of apoptotic, necrotic and healthy cells within the same cell population by flow cytometry or fluorescence microscopy.
1. Apoptotic cell detection - green
During apoptosis, phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. Using Annexin V, which has a high affinity for PS, labelled with CF™488, this kit identifies apoptotic cells highlighting them with green fluorescence by binding to PS exposed on the outer leaflet. CF™488A is spectrally similar to fluorescein (FITC) but offers a much brighter and more photostable fluorescence.
2. Necrotic cell detection - red
During necrosis, both internal organelle and plasma membrane integrity are lost, resulting in spilling of cytosolic and cellular contents into the surrounding environment. Utilising Ethidium homodimer III (EtD-III), a highly positively charged nucleic acid probe which is impermeant to live cells or apoptotic cells, this kit stains and identifies necrotic cells with red fluorescence (λabs/λem = 528/617 nm). EthD-III is a superior alternative to propidium iodide (PI) or ethidium homodimer I used in other kits due to its significantly higher affinity for DNA and higher fluorescence quantum yield.
3. Healthy cells detection - blue
Utilising Hoechst 33342, a cell membrane-permeant, minor groove-binding DNA stain, this kit identifies healthy cells by emitting bright blue fluorescence upon binding to DNA (λabs/λem = 350/461 nm). Hoechst 33342 stains the nuclei of all cells blue. Healthy cells can be differentiated from apoptotic/necrotic cells as the former healthy cells will be stained by Hoechst only, and the latter will be stained both blue and green (CF™488-Annexin V) and blue and red (EtD-III) respectively.
Benefits of using the Apoptotic, Necrotic and Healthy Cell Quantification Kit Plus
• Simultaneous detection of apoptotic, necrotic and healthy cells in a single assay
• Simple to use
• Single 15-30 minute staining step
• Sensitive - EthD-III is a superior alternative to both PI and EthD-I for staining necrotic cells
• Uses the photostable and bright CF488™ dye
• Analysis by flow cytometry or fluorescence microscopy
Material available for download
Apoptosis, Necrosis and Healthy Cell Quantitation Kit Plus protocol
Note: product availability depends on country - see product detail page.
|Details||Cat number & supplier||Size||Price|
|Apoptotic, Necrotic & Healthy Cells Quantitation Kit Plus (50 assays) BT30066 · Biotium||