SR-FLICA® Poly Caspase Assay Kit

$195.00$561.00

Detect active caspase enzymes with the SR FLICA® Poly Caspase Assay Kit. This in vitro assay employs the fluorescent inhibitor probe SR-VAD-FMK to label active caspase enzymes in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.

Catalog # Size Quantity Price
916 25 Tests $195.00
917 100 Tests $561.00
Catalog #: 916, 917 Categories: , ,

Caspases play important roles in apoptosis and inflammation. ICT’s FLICA poly caspase assay kit is used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells and tissues.

The FLICA reagent SR-VAD-FMK enters each cell and irreversibly binds to activated caspases. Because the SR-VAD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound SR-VAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining red 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 Hoechst 33342 to detect changes in nuclear morphology.

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 – 1:60. For example, to stain at 1:30, add 10 μL to 290 μL of cultured cells. To stain at 1:60, add 5 μL to 295 μL of cultured cells.
6. Incubate approximately 1 hour.
7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buff er and spin cells.
8. If desired, label with additional stains, such as Hoechst, 7-AAD, or an antibody.
9. If desired, fix or embed cells.
10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. SR-FLICA excites at 550-580 nm and emits at 590-600 nm.

Jurkat cells were treated with 1 µM staurosporine to induce caspase activity (top), or a negative control (bottom) for 3 hours, incubated with ICT’s red poly caspases inhibitor probe, SR-VAD-FMK, for 1 hour, washed twice, and examined under a fluorescence microscope (DIC images were also taken). The color image of induced cells (upper left) reveals experimental cells which fluoresce red, therefore they all have some degree of caspase activity. The non-induced DIC image (lower right) reveals many control cells, however the corresponding fluorescence image (lower left) is dark; none of these cells have active caspases (Dr. Brian W. Lee, ICT).

srvaddata

• Reagent name: SR-VAD-FMK
• Target: Poly caspases
• Excitation/Emission: 550-580 nm / 590-600 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

Kit 916: 25 Tests
• FLICA Poly Caspase Reagent (SR-VAD-FMK), 1 vial, #679
• 10X Apoptosis Wash Buffer, 15 mL, #635
• Fixative, 6 mL, #636
• Hoechst 33342, 1 mL, #639
• Kit Manual

Kit 917: 100 Tests
• FLICA Poly Caspase Reagent (SR-VAD-FMK), 4 vials, #679
• 10X Apoptosis Wash Buffer, 60 mL, #634
• Fixative, 6 mL, #636
• Hoechst 33342, 1 mL, #639
• Kit Manual

Nougarede A, Popgeorgiev N, Kassem L, Omarjee S, Borel S, Mikaelian I, Lopez J, Gadet R, Marcillat O, Treilleux I, Villoutreix BO, Rimokh R, Gillet G. Breast cancer targeting through inhibition of the endoplasmic reticulum-based apoptosis regulator Nrh/BCL2L10. Cancer Res. 2018. Jan 12. pii: canres.0846.2017. doi: 10.1158/0008-5472.CAN-17-0846. [Epub ahead of print]. Abstract

“Apoptosis was assessed at the end of the corresponding drug treatment using the SR-FLICA® Poly Caspase Assay staining (ImmunoChemistry Technologies). Images were acquired using a Nikon NiE microscope and green transfected cells positive for FLICA staining were counted with a custom- made …”

Schell SL, Soni C, Fasnacht MJ, Domeier PP, Cooper TK, Rahman ZSM. Mer Receptor Tyrosine Kinase Signaling Prevents Self-Ligand Sensing and Aberrant Selection in Germinal Centers. J. Immunol. Dec 15;199(12):4001-4015. doi: 10.4049/jimmunol.1700611. Epub 2017 Nov 8. Abstract

” To detect AC accumulation, the SR-FLICA Poly Caspase Assay kit was purchased from ImmunoChemistry Technologies and the protocol was performed according to the manufacturer’s instructions.”

You may also like…