Testing Data
(Jurkat suspension cells were exposed to 1 uM of staurosporine for 4 hours at 37 degree C to induce apoptosis. Cells were dually stained with the green fluorescent FAM-FLICA poly caspase probe to detect apoptosis via caspase activity, and the red fluorescent vital dye 7-AAD to detect necrosis. The image shown in panel A reveals 4 populations of cells: 1) Live, unstained cells, which do not fluoresce. 2) Early stage apoptotic cells fluoresce green with FAM-FLICA. 3) Dually stained green and red fluorescing cells represent the population of Jurkat cells in mid-to-late stage apoptosis; these cells have active caspase enzymes and compromised cell membranes. 4) Necrotic cells fluoresce red. Panel B shows a corresponding differential interference contrast (DIC) image, which reveals cell morphology.)
Necrosis vs Apoptosis Assay Kit
Necrosis vs Apoptosis Assay Kit
Testing Data
(Jurkat suspension cells were exposed to 1 uM of staurosporine for 4 hours at 37 degree C to induce apoptosis. Cells were dually stained with the green fluorescent FAM-FLICA poly caspase probe to detect apoptosis via caspase activity, and the red fluorescent vital dye 7-AAD to detect necrosis. The image shown in panel A reveals 4 populations of cells: 1) Live, unstained cells, which do not fluoresce. 2) Early stage apoptotic cells fluoresce green with FAM-FLICA. 3) Dually stained green and red fluorescing cells represent the population of Jurkat cells in mid-to-late stage apoptosis; these cells have active caspase enzymes and compromised cell membranes. 4) Necrotic cells fluoresce red. Panel B shows a corresponding differential interference contrast (DIC) image, which reveals cell morphology.)
Testing Data
(Jurkat suspension cells were exposed to 1 uM of staurosporine for 4 hours at 37 degree C to induce apoptosis. Cells were dually stained with the green fluorescent FAM-FLICA poly caspase probe to detect apoptosis via caspase activity, and the red fluorescent vital dye 7-AAD to detect necrosis. The image shown in panel A reveals 4 populations of cells: 1) Live, unstained cells, which do not fluoresce. 2) Early stage apoptotic cells fluoresce green with FAM-FLICA. 3) Dually stained green and red fluorescing cells represent the population of Jurkat cells in mid-to-late stage apoptosis; these cells have active caspase enzymes and compromised cell membranes. 4) Necrotic cells fluoresce red. Panel B shows a corresponding differential interference contrast (DIC) image, which reveals cell morphology.)
Testing Data
( Flow cytometry analysis of Jurkat suspension cells to quantify four populations. (A) Cells were treated with a placebo (non-induced treatment with DMSO). (B) Cells were treated with 1 uM staurosporine for 4 hours to induce apoptosis via caspase activity. Cells were then dually stained with FAM-FLICA and 7-AAD, and analyzed using an Accuri C6 flow cytometer. FAM-FLICA was analyzed on FL-1 and 7-AAD was analyzed on FL-3. The key is shown in (C). Live, unstained cells do not fluoresce (lower left quadrant). Early apoptotic cells fluoresce green with FAM-FLICA. Dually stained green and red fluorescing cells represent the population of cells in mid to late apoptosis (these cells have active caspase enzymes and compromised cell membranes). Necrotic cells fluoresce red. In the non-induced population (A), only 9.8% of cells were apoptotic (LR: 4.5% + UR: 6.3%), compared with 97.1% of the induced population (B; LR: 46.4% + UR: 50.7%).)
Testing Data
( Flow cytometry analysis of Jurkat suspension cells to quantify four populations. (A) Cells were treated with a placebo (non-induced treatment with DMSO). (B) Cells were treated with 1 uM staurosporine for 4 hours to induce apoptosis via caspase activity. Cells were then dually stained with FAM-FLICA and 7-AAD, and analyzed using an Accuri C6 flow cytometer. FAM-FLICA was analyzed on FL-1 and 7-AAD was analyzed on FL-3. The key is shown in (C). Live, unstained cells do not fluoresce (lower left quadrant). Early apoptotic cells fluoresce green with FAM-FLICA. Dually stained green and red fluorescing cells represent the population of cells in mid to late apoptosis (these cells have active caspase enzymes and compromised cell membranes). Necrotic cells fluoresce red. In the non-induced population (A), only 9.8% of cells were apoptotic (LR: 4.5% + UR: 6.3%), compared with 97.1% of the induced population (B; LR: 46.4% + UR: 50.7%).)
Background: Assessing potential cytotoxicity properties of chemical and biological agents is a mandatory requirement for the safe distribution of pharmaceuticals, vaccines, or additives associated with food product formulations. The Necrosis vs Apoptosis Assay kit simultaneously detects both apoptosis associated cytotoxicity events as well as cell death due to necrosis. Apoptotic cells are identified using the FLICA reagent probe. The FAM-FLICA probe covalently binds to active caspase enzymes, which are up-regulated during apoptosis, thus clearly labeling apoptotic cells for subsequent analysis. Non-apoptotic cells will not contain the active caspase enzymes required for FAM-FLICA to remain covalently bound within the cell structure. Loss of the integrity of the cell membrane, indicative of necrosis or late stage apoptosis, is detected using the vital staining dye, 7-aminoactinomycin D (7-AAD), a red fluorescing live/dead stain. This dye easily penetrates cell membrane-compromised cells, binding tightly to GC rich regions of the DNA. Because 7-AAD alone may not detect cells in the early stages of apoptosis, it is essential to use it in combination with the green-fluorescent FAM-FLICA apoptosis detection reagent. Combining these two different types of fluorescent cell-status-indicator reagents within a single test can reveal a significant percentage of cells that are 7-AAD-negative (membrane intact live cells) and yet FAM-FLICA positive (apoptotic)
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Product Notes
The Necrosis vs Apoptosis (Catalog #AAA258047) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Necrosis vs Apoptosis, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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