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Invitrogen™ Vybrant™ Apoptosis Assay Kits for apoptotic and necrotic cell staining
Description
Apoptotic cells undergo several physiological processes, including changes in cell membrane permeability and chromatin condensation, which can be distinguished using different dyes in apoptotic and necrotic cell staining protocols. Vybrant Apoptosis Assay kits offer streptavidin biotin-annexin V, propidium iodide (PI), YO-PRO-1, and Hoechst 33342 dyes to differentiate live from apoptotic or necrotic cells, which can then be visualized using flow cytometry.
The Vybrant Apoptosis Assay Kit #4 detects apoptosis via changes that occur in cell membrane permeability. This kit contains ready-to-use solutions of both YO-PRO-1 and propidium iodide (PI) nucleic acid stains. YO-PRO-1 stain selectively passes through the plasma membranes of apoptotic cells and labels them with moderate green fluorescence. Necrotic cells are stained fluorescent red with PI.
The Vybrant Apoptosis Assay Kit #5, or the Chromatin Condensation/Dead Cell Apoptosis Kit with Hoechst 33342 and PI for Flow Cytometry, provides a rapid and convenient assay for apoptosis based on fluorescence detection of the compacted state of the chromatin in apoptotic cells. This kit contains ready-to-use solutions of blue-fluorescent Hoechst 33342 dye, which stains the condensed chromatin of apoptotic cells more brightly than the chromatin of non-apoptotic cells, and red-fluorescent PI dye, which stains dead cells.
The Vybrant Apoptosis Assay Kit #6 provides a rapid and convenient assay for apoptosis, using the bright, blue-fluorescent Alexa Fluor 350 conjugate of streptavidin in conjunction with biotin-X annexin V to detect apoptotic cell populations either by flow cytometry or imaging. In addition, the kit includes a ready-to-use solution of the fluorescent red PI nucleic acid–binding dye. PI is impermeant to live cells and apoptotic cells, but stains dead cells with red fluorescence, binding tightly to their nucleic acid. The kit contains an ample amount of each reagent for about 50 flow cytometric assays.
Specifications
Specifications
| Description | Vybrant Apoptosis Assay Kit #6 |
| Quantity | 50 assays |
| Format | Tube |
| Product Type | Apoptosis Kit |
| Content And Storage | 1 vial of biotin-x annexin V, 1 vial of Alexa Fluor™ 350 streptavidin, 1 vial of propidium iodide, and 1 bottle of 5x annexin binding buffer. Store in refrigerator (2–8°C) and protect from light |
| Flow Cytometer Laser Lines | UV, 488 |
| Excitation/Emission | Alexa Fluor™ 346/442: 346 PI: 535/617 |
| Color | Blue, Red |
| Shipping Condition | Wet Ice |
| Product Line | Alexa Fluor |
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Frequently Asked Questions (FAQs)
YO-PRO-1: 491/509 nm, bound to DNA
Propidium iodide: 535/617 nm, bound to DNA
See Table 1 in this article (http://www.thermofisher.com/content/dam/LifeTech/migration/en/filelibrary/support/bioprobes/bioprobes-65.par.61751.file.dat/bioprobes-65-cellevent.pdf) where a Jurkat model system was induced with 10 µM camptothecin for time periods of 0 to 4 hours. It is important to note that these results were studied using a single cell type and induction system; results may differ for other experimental systems. Increased membrane permeability in apoptotic cells can be discriminated from dead cells using YO-PRO-1 dye in combination with propidium iodide or the SYTOX dead cell indicators. Other mid-apoptotic events are increased ROS production (detected with CellROX reagents, H2DCFDA), changes in cellular pH (BCECF, SNARF-1) and calcium release (Fluo-4, Fura-2, Indo-1). No single parameter defines apoptosis under any condition, so it is best to employ a multi-parametric approach when studying apoptosis.