Article No
260-92
FAM-FLICA® Poly Caspase Assay Kit
FAM-FLICA® Poly Caspase Assay Kit
Assay & Detection
Specifications
| Application | Activity Assay |
| Article No | 260-92 |
| Country Availability | SE, FI, DK, NO, IS, EE, LV, LT |
| Components | Kit 92: 100 tests • FLICA Poly Caspase Reagent (FAM-VAD-FMK), 4 vials, #637 • 10X Apoptosis Wash Buffer, 60 mL, #634 • Fixative, 6 mL, #636 • Propidium Iodide, 1 mL, #638 • Hoechst 33342, 1 mL, #639 • Kit Manual |
| Description | FAM-FLICA® Poly Caspase Assay Kit |
| Supplier | ImmunoChemistry Technologies |
| Notes | Caspases play important roles in apoptosis and inflammation. ICT’s FLICA assay kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells and tissues. The FAM FLICA Poly Caspase probe allows researchers to assess caspase activation. The FLICA reagent FAM-VAD-FMK enters each cell and irreversibly binds to activated caspases. Because the FAM-VAD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound FAM-VAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of the active caspase enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by a fluorescence plate reader, fluorescence microscopy or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative included in the kit. Unfixed samples may also be analyzed with Propidium Iodide or Hoechst 33342 to detect changes in necrosis or nuclear morphology, respectively. |
| Product Type | Assay & Detection |
| Protocol | 1. Prepare samples and controls 2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH20. 3. Reconstitute FLICA 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 to 290 µL of cultured cells). 6. Incubate approximately 1 hour. 7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells. 8. If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody. 9. If desired, fix cells. 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLICA excites at 492 nm and emits at 520 nm. If working with adherent cells, please see the manual for additional protocols. |
| Shipping Information | BLUE ICE |
| Size | 100 Tests |
| Storage | 4°C |
| Technical Specifications | • Reagent name: FAM-VAD-FMK • Target: Poly caspases • Excitation/Emission: 492 nm / 520 nm • Method of Analysis: Flow Cytometer, Fluorescence Plate Reader, Fluorescence Microscope • Types of Samples: Cell culture • Storage: 2-8°C • Shipping: Ships overnight (domestic), International Priority Shipping |
| Product Page Updated | 2024-01-03T12:33:39.983Z |