Product Details
Catalog Number: CT17004
Type: Kit
Storage: Store at 4°C-8°C.
Shipping: Refrigerated (Polar Packs)
Product Sizes
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250 Tests$585.00Add to Cart

Kit Specifications

 

Target:Cell-mediated cytotoxicity and apoptosis (poly caspases)
Excitation / Emission:SR FLICA: 565 nm / 590 nm · CFSE: 488 nm / 520 nm · 7-AAD: 545 nm / 650 nm
Method of Analysis:Flow Cytometer
Kit Contents:
  • FLICA Poly Caspases Reagent (SR-VAD-FMK), 1 or 2 vials
  • CFSE target cell stain, 1 vial
  • 7-AAD vital stain, 1 or 2 vials
  • 10x Assay Buffer, 30 or 60 mL

Test Kit Steps:

  1. Dilute 10X assay buffer in DI water and filter to sterilize.
  2. Prepare required controls.
  3. Make single-cell preparation of target cell population with final resuspension in assay buffer at 10^5 - 10^6 cells/ml.
  4. Reconstitute CFSE in DMSO and dilute with assay buffer to working concentration.
  5. Label target cells with CFSE for 15 min at room temperature.
  6. Wash unbound label from target cells (may eliminate step if no effector cells are to be added).
  7. Adjust target cells to appropriate concentration and add either (a) effector cells to obtain desired E/T ratios, or (b) desired drug or antibody + complement to induce killing.
  8. Incubate at 37°C for desired time period to induce killing.
  9. Reconstitute SR-FLICA with DMSO and dilute in assay buffer to working concentration.
  10. Add SR-FLICA reagent and incubate at 37°C for 45 min.
  11. Reconstitute 7-AAD with DMSO and dilute in sterile assay buffer. 1
  12. Add 7-AAD to cells and incubate on ice for 10 min. 1
  13. Set up cytometer and analyze samples. 
Images

All target cells are stained green with CFSE (R3 = R1+R2): live target cells are in the lower right quadrant (R1); necrotic and late apoptotic target cells are dually stained red with 7-AAD and are shown in the upper right quadrant (R2).

All target cells are stained green with CFSE (R3 = R1+R2): live target cells are in the lower right quadrant (R1); necrotic and late apoptotic target cells are dually stained red with 7-AAD and are shown in the upper right quadrant (R2).