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Annexin V-FITC Apoptosis Detection Kit with SYTOX (ab14086) includes Annexin V-FITC, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-FITC and SYTOX Green dye in the provided binding buffer, apoptotic cells show green fluorescence, dead cells show a higher level of green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished using a flow cytometry with the 488 nm line of an argon-ion laser for excitation. Both annexin V-FITC and SYTOX Green dye emit green fluorescence that can be detected in the FL1 channel, freeing the other channels for the addition of other probes in multi-color labeling experiments.
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The assay is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells, without the need of fixation.
|Annexin V-FITC||1 x 500µl|
|Binding Buffer||1 x 50ml|
|SYTOX Green Dye||1 x 100µl|
Our Abpromise guarantee covers the use of ab14086 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent dilution.|
|FM||Use at an assay dependent concentration.|
ab14086 has not yet been referenced specifically in any publications.