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If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
The binding buffer included in the kit needs to be used for resuspending cells. The buffer contains calcium chloride at a final (1X) concentration of 2.5 mM which is necessary for the binding of annexin V to phosphatidylserine.
Annexin V FITC will have a maximum emission of 528 nm. This can be measured in the standard FITC Channel on a flow cytometer (FL1). Propidium Iodide has a maximum emission of 620 nm. This is measured on the short red channel on a flow cytometer (FL2 or FL3).
When using a kinetic study (various time points), you can show the progression of the cells from viable (annexin V FITC negative, PI negative), to annexin FITC positive, PI negative (membrane flip) to annexin V FITC positive, PI positive (dead). If there are cells that are double positive when starting, it is not possible to guarantee that the cell death occurred due to apoptosis with this assay.
It is not possible to have cells that are PI positive without the cells also being positive for Annexin V binding. Cells are PI positive because the membrane has been compromised. If this is the case, Annexin V can also enter the cell and bind to the PS on the internal cell membrane. The gating on the histogram for the FITC channel should be changed so that all cells that are PI positive are also Annexin V positive.
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
No. Product APOAF, Annexin V-FITC Apoptosis Detection Kit must be performed on live cells in order to measure Apoptosis. The assay is based on the externization of phosphatidylserine from the inner cell membrane to the outer cell membrane. If the membrane is preturbed due to fixation, non-speciifc staining of the inner cell membrane might occur
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