SR-FLICA® Caspase-3/7 Assay Kit

$195.00$561.00

Detect caspase 3 and 7 activity with the SR-FLICA® Caspase-3/7 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe SR-DEVD-FMK to label active caspase 3 and 7 enzymes in living cells. Analyze the fluorescent signal using fluorescence microscopy, a fluorescence plate reader, or by flow cytometry.

Catalog # Size Quantity Price
931 25 Tests $195.00
932 100 Tests $561.00
Catalog #: 931, 932 Categories: , ,

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 FLICA reagent SR-DEVD-FMK enters each cell and irreversibly binds to activated caspase 3 and 7. Because the SR-DEVD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound SR-DEVD-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 3 and 7 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. Unfixed samples may also be analyzed with Hoechst 33342 to detect changes in nuclear morphology.

1. Prepare samples and controls.
2. Dilute 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 Buffer 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.

Mouse EL4 thymoma cells were cultured in the absence (A) or presence (B) of 100 µg mitomycin for 18 hours. The cells were harvested and stained with ICT’s SR-DEVD-FMK inhibitor probe (red histograms) to detect active caspases-3/7. The data illustrate a low level of caspase-3 activity in untreated cells (A) compared to unstained control (blue histogram); caspase-3 activity is greatly enhanced in the mitomycin treated cultures (B). Cells were analyzed with a BD LSRII flow cytometer to detect sulforhodamine using the green laser (532 nm) and detector D set up with filters 610/20 and 600 LP (Dr. Catherine Martone, Yale University).

srdevddata

• Reagent name: SR-DEVD-FMK
• Target: Caspase-3/7
• Excitation/Emission: 550-580 nm / 590-600 nm
• Method of Analysis: Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
• Types of Samples: Cell culture
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping

Kit 931: 25 Tests
• FLICA Caspase-3/7 Reagent (SR-DEVD-FMK), 1 vial, #678
• 10X Apoptosis Wash Buffer, 15 mL, #635
• Fixative, 6 mL, #636
• Hoechst 33342, 1 mL, #639
• Kit Manual

Kit 932: 100 Tests
• FLICA Caspase-3/7 Reagent (SR-DEVD-FMK), 4 vials, #678
• 10X Apoptosis Wash Buffer, 60 mL, #634
• Fixative, 6 mL, #636
• Hoechst 33342, 1 mL, #639
• Kit Manual

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