SPONTANEOUS APOPTOSIS ASSAY V. 2.0

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Tony Rook
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SPONTANEOUS APOPTOSIS ASSAY V. 2.0

Please find the link for the following protocol:

SPONTANEOUS APOPTOSIS ASSAY V. 2.0

http://aactg.s-3.com/pub/download/im.0popto.pdf

Principles and Purpose of the Test:

Peripheral blood mononuclear cells (PBMC) isolated from HIV-1-infected persons undergospontaneous apoptosis upon in-vitro cultivation. The method using propidium iodide staining (PI) and measurement of DNA content from PBMC isolated from HIV-1-infected persons to determine percentage apoptosis relies on the fact that PI will stoichiometrically stain cellular DNA following methanol fixation and RNase A treatment. Apoptotic cells have more fragmented DNA, which will be lost from the cell following the methanol fixation step. Thus, after staining with PI, these apoptotic cells will have a lower DNA content than non-apoptotic cells and will appear as a peak or smear with less fluorescence. Reduction in the value of percent apoptosis in PBMC obtained from HIV-1-infected persons over a time period after treatment with certain agents can aid to evaluate the effect of therapy and the progression of the disease.