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Cell Meter™ PE-Annexin V Binding Apoptosis Assay Kit
Optimized for Flow Cytometry
Annexin V may be conjugated to fluorochromes including PE. This format retains its high affinity for phosphatidylserine (PS) and thus serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. Since externalization of PS occurs in the earlier stages of apoptosis, PE Annexin V staining can identify apoptosis at an earlier stage than assays based on nuclear changes such as DNA fragmentation. PE Annexin V staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from either apoptotic or necrotic processes. Therefore, staining with PE Annexin V is typically used in conjunction with a vital dye such as propidium iodide (PI) or 7-Amino-Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells (7-AAD negative, PE Annexin V positive). Viable cells with intact membranes exclude 7-AAD, whereas the membranes of dead and damaged cells are permeable to 7-AAD. For example, cells that are considered viable are both PE Annexin V and 7-AAD negative while cells that are in early apoptosis are PE Annexin V positive and 7-AAD negative, while cells that are in late apoptosis or already dead are both PE Annexin V and 7-AAD positive. This assay does not distinguish between cells that have undergone apoptotic death versus those that have died as a result of a necrotic pathway because in either case, the dead cells will stain with both PE Annexin V and 7-AAD. However, when apoptosis is measured over time, cells can be often tracked from PE Annexin V and 7-AAD negative (viable, or no measurable apoptosis), to PE Annexin V positive and 7-AAD negative (early apoptosis, membrane integrity is present) and finally to PE Annexin V and 7-AAD positive (end stage apoptosis and death). The movement of cells through these three stages suggests apoptosis. In contrast, a single observation indicating that cells are both PE Annexin V and 7-AAD positive, in of itself, reveals less information about the process by which the cells underwent their demise.
The detection of binding activity of RPE-Annexin V to phosphatidylserine in Jurkat cells with Cell Meter™ RPE-Annexin V Binding Apoptosis Assay Kit. Jurkat cells were treated without (Blue) or with 1 µM staurosporine (Red) in a 37 °C, 5% CO2 incubator for 4-5 hours, and then dye loaded with RPE-Annexin V for 30 minutes. The fluorescence intensity of RPE-Annexin V was measured with a FACSCalibur (Becton Dickinson) flow cytometer using the FL2 channel.
The detection of binding activity of RPE-Annexin V to phosphatidylserine in Jurkat cells with Cell Meter™ RPE-Annexin V Binding Apoptosis Assay Kit. Jurkat cells were treated without (Blue) or with 1 µM staurosporine (Red) in a 37 °C, 5% CO2 incubator for 4-5 hours, and then dye loaded with RPE-Annexin V for 30 minutes. The fluorescence intensity of RPE-Annexin V was measured with a FACSCalibur (Becton Dickinson) flow cytometer using the FL2 channel.
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
22838100 Tests
Price
 
Spectral properties
Correction factor (280 nm)0.175
Extinction coefficient (cm -1 M -1)
1960000
Excitation (nm)565
Emission (nm)574
Quantum yield
0.82
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Flow cytometer
Excitation488 nm or 561 nm laser
Emission575/26 nm filter
Instrument specification(s)PE channel
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Page updated on October 8, 2024