Necrosis vs Apoptosis Assay Kit

This kit simultaneously detects both cell death due to both apoptosis and necrosis. It can be used to assess the effects of novel therapeutic agents on cell health. Analyze the fluorescent signal using fluorescence microscopy or flow cytometry.

Figure 1. Using FAM-FLICA®, non-apoptotic cells (unstained, left of histogram) can easily be distinguished from apoptotic cells exhibiting green fluorescence (right of histogram). Jurkat cells were either treated with DMSO as control (A), or 1 µM staurosporine (B) to induce apoptosis. Cells were stained with FAM-FLICA® and analyzed using a flow cytometer in FL-1. Only 7% of non-induced cells (A, right) are apoptotic compared with 96.5% of the induced cells (B, right). (ICT 226:17-19.)
Figure 2. Using 7-AAD, live cells (unstained, left side of each histogram) can easily be distinguished from dead/membrane compromised cells exhibiting red fluorescence (right side of each histogram). Jurkat cells were either untreated (A), or treated with 90% ethanol for 60 seconds, then stained with 7-AAD, and analyzed using a flow cytometer in FL-3. Only 4% of untreated cells (A) are dead compared with 97.6% of the treated cells (B). Data courtesy of Dr. Kristi Strandberg, ICT 226:30-31.
Figure 3. Jurkat suspension cells were exposed to staurosporine, and then stained with FAM-FLICA® and 7-AAD. Cells in early stage apoptosis fluoresce green with FAM-FLICA®, cells in mid-to-late stage apoptosis fluoresce green with FAM-FLICA® and red with 7-AAD, and necrotic cells fluoresce red with 7-AAD. Panel B the corresponding DIC image. FAM-FLICA® and 7-AAD were imaged using a 470-490 nm excitation filter plus >520 nm long pass filter tandem. Data courtesy of Dr. Brian W. Lee (ICT 196:70).
Figure 4. Live, unstained cells do not fluoresce (LL, lower left). Early stage apoptotic cells fluoresce green with FAM-FLICA® (LR, lower right). Dually stained green and red fluorescing cells represent cells in mid to late stage apoptosis; these cells have active caspase enzymes and compromised cell membranes (UR, upper right). Necrotic cells fluoresce red (UL, upper left).
Figure 5. Jurkat cells were treated with DMSO or staurosporine to induce apoptosis, and then stained with FAM-FLICA® (FL-1) and 7-AAD (FL-3). Live cells (lower left, B and D). Early stage apoptotic cells (lower right, B and D). Mid to late stage apoptosis (upper right, B and D). Necrotic cells (upper left, B and D). See Figure 4 for key. Only 10.8% of non-induced cells were apoptotic (LR: 4.5% + UR: 6.3%), compared with 97.1% of induced cells (D; LR: 46.4% + UR: 50.7%). ICT 226:17-19).
Figure 6. U-2 OS cells were treated with a placebo or staurosporine to induce apoptosis, and then stained with FAM-FLICA® (FL-1) and 7-AAD (FL-3). Live cells (lower left, B and D). Early stage apoptotic cells (lower right, B and D). Mid to late stage apoptosis (upper right, B and D). Necrotic (upper left, B and D). See Figure 4 for key. Only 5.9% of non-induced cells were apoptotic (LR: 3.6% + UR: 2.3%) compared with 30.5% of induced cells (D; LR: 22.1% + UR: 8.4%). (ICT 226:17-19).
Figure 1. Using FAM-FLICA®, non-apoptotic cells (unstained, left of histogram) can easily be distinguished from apoptotic cells exhibiting green fluorescence (right of histogram). Jurkat cells were either treated with DMSO as control (A), or 1 µM staurosporine (B) to induce apoptosis. Cells were stained with FAM-FLICA® and analyzed using a flow cytometer in FL-1. Only 7% of non-induced cells (A, right) are apoptotic compared with 96.5% of the induced cells (B, right). (ICT 226:17-19.)
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Figure 1. Using FAM-FLICA®, non-apoptotic cells (unstained, left of histogram) can easily be distinguished from apoptotic cells exhibiting green fluorescence (right of histogram). Jurkat cells were either treated with DMSO as control (A), or 1 µM staurosporine (B) to induce apoptosis. Cells were stained with FAM-FLICA® and analyzed using a flow cytometer in FL-1. Only 7% of non-induced cells (A, right) are apoptotic compared with 96.5% of the induced cells (B, right). (ICT 226:17-19.)
Figure 2. Using 7-AAD, live cells (unstained, left side of each histogram) can easily be distinguished from dead/membrane compromised cells exhibiting red fluorescence (right side of each histogram). Jurkat cells were either untreated (A), or treated with 90% ethanol for 60 seconds, then stained with 7-AAD, and analyzed using a flow cytometer in FL-3. Only 4% of untreated cells (A) are dead compared with 97.6% of the treated cells (B). Data courtesy of Dr. Kristi Strandberg, ICT 226:30-31.
Figure 3. Jurkat suspension cells were exposed to staurosporine, and then stained with FAM-FLICA® and 7-AAD. Cells in early stage apoptosis fluoresce green with FAM-FLICA®, cells in mid-to-late stage apoptosis fluoresce green with FAM-FLICA® and red with 7-AAD, and necrotic cells fluoresce red with 7-AAD. Panel B the corresponding DIC image. FAM-FLICA® and 7-AAD were imaged using a 470-490 nm excitation filter plus >520 nm long pass filter tandem. Data courtesy of Dr. Brian W. Lee (ICT 196:70).
Figure 4. Live, unstained cells do not fluoresce (LL, lower left). Early stage apoptotic cells fluoresce green with FAM-FLICA® (LR, lower right). Dually stained green and red fluorescing cells represent cells in mid to late stage apoptosis; these cells have active caspase enzymes and compromised cell membranes (UR, upper right). Necrotic cells fluoresce red (UL, upper left).
Figure 5. Jurkat cells were treated with DMSO or staurosporine to induce apoptosis, and then stained with FAM-FLICA® (FL-1) and 7-AAD (FL-3). Live cells (lower left, B and D). Early stage apoptotic cells (lower right, B and D). Mid to late stage apoptosis (upper right, B and D). Necrotic cells (upper left, B and D). See Figure 4 for key. Only 10.8% of non-induced cells were apoptotic (LR: 4.5% + UR: 6.3%), compared with 97.1% of induced cells (D; LR: 46.4% + UR: 50.7%). ICT 226:17-19).
Figure 6. U-2 OS cells were treated with a placebo or staurosporine to induce apoptosis, and then stained with FAM-FLICA® (FL-1) and 7-AAD (FL-3). Live cells (lower left, B and D). Early stage apoptotic cells (lower right, B and D). Mid to late stage apoptosis (upper right, B and D). Necrotic (upper left, B and D). See Figure 4 for key. Only 5.9% of non-induced cells were apoptotic (LR: 3.6% + UR: 2.3%) compared with 30.5% of induced cells (D; LR: 22.1% + UR: 8.4%). (ICT 226:17-19).

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Product Manual
Safety Data Sheet

SKU: 9148

Size: 100 - 200 tests
Price:
$641.00
Ships: 1-2 business days
  1. Prepare samples and controls
  2. Dilute Apoptosis Wash Buffer 1:10 with diH2O.
  3. Reconstitute FAM-VAD-FMK with 50 µL DMSO.
  4. Dilute FLICA 1:5 by adding 200 µL PBS.
  5. Add diluted FLICA to each sample at 1:30 (e.g., add 10 µL of diluted FLICA to 290 µL of sample).
  6. Incubate approximately 1 hour.
  7. Remove media, then wash cells: add 1X Apoptosis Wash Buffer and spin cells (twice).
  8. Resuspend samples in 400 µL 1X Apoptosis Wash Buffer
  9. Reconstitute 7-AAD with 260 µL DMSO
  10. Add 7-AAD to each sample at 1:200 (e.g., add 2 µL to 400 µL of sample)
  11. Analyze with a fluorescence microscope or flow cytometer. FAM-VAD-FMK excites at 492 nm and emits at 529 nm. 7-AAD excites at 546 nm and emits at 647 nm.
Kit 9147: 50-100 Tests
  • FAM-FLICA Poly Caspase Reagent (FAM-VAD-FMK), 2 vials, #637
  • 7-Aminoactinomycin D (7-AAD) vital dye, 1 0.26 mg vial, #6163
  • 10X Apoptosis Wash Buffer, 1 60 mL bottle, #634
  • Fixative, 6 mL, #636
  • Kit Manual
    • Kit 9148: 100-200 Tests
  • FAM-FLICA Poly Caspase Reagent (FAM-VAD-FMK), 4 vials, #637
  • 7-Aminoactinomycin D (7-AAD) vital dye, 2 0.26 mg vials, #6163
  • 10X Apoptosis Wash Buffer, 2 60 mL bottles, #634
  • Fixative, 6 mL, #636
  • Kit Manual

    • Bulk Order Necrosis vs Apoptosis Assay Kit

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