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Serine Protease Assay Kit

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Active chymotrypsin-like serine protease activity staining by flow cytometry. Jurkat cells were treated as shown with 1mM staurosporine to induce apoptosis. The cells were then stained using the FLISP FAM-Phe-DAP Serine Protease Assay Kit including propidium iodide (APO009). Apoptotic cells positive for serine protease activity can be seen in the bottom right quadrant and dead cells positive for both serine protease and PI in the top right quadrant. Healthy cells are negative for both stains. Data acquired on the ZE5™ Cell Analyzer.

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FLISP™ FAM-Phe-DAP Serine Protease Assay Kit

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Serine Protease Assay Kit

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Product Code Applications Pack Size List Price Your Price Qty
APO009
Datasheet Datasheet Datasheet
SDS Safety Datasheet SDS
F IF 25 Tests
List Price Your Price
Search for Batch Specific Datasheets

Fluorescent Labeled Inhibitors of Serine Protease (FLISP) kits allow the analysis of intracellular chymotrypsin-like serine protease activity in live cells. Serine protease activation plays major roles in several different cellular functions including apoptosis, tumor malignancy and as diagnostic and prognostic indicators of breast, head and neck carcinomas. Serine proteases are also thought to play a role in infections and transplant rejection.

Reagents in the Kit
FAM-Phe-DAP FLISP Reagent - lyophilized
10X Wash Buffer, 15 mL
Fixative, 6 mL
Propidium Iodide, 1 mL
Hoechst Stain, 1 mL
Test Principle
Active chymotrypsin-like enzymes within the cell bind covalently to the FLISP FAM-Phe-DAP inhibitor probe, retaining the green fluorescent signal within the cell. The higher the concentration of active chymotrypsin-like enzymes within the cell, the higher the level of fluorescence observed. FLISP kits are suitable for use with suspension or adherent cells and can be used with a flow cytometer, fluorescence microscope or a fluorescence plate reader.

FLISP inhibitors are cell-permeable and non-cytotoxic fluorochrome-labeled analogs of the first serine protease inhibitor, tosyl-phenlalanylchloromethyl ketone (TPCK), labeled with carboxyfluorescein (FAM) which fluoresces green. They are available with either a chloromethyl ketone (CMK) or diphenyl 1-(N-peptidylamino) alkanephosphonate (DAP) reactive group containing compound. Specificity for chymotrypsin-like enzymes is determined by the amino acid targeting peptide, the FAM-Phe-DAP probe inhibits chymotrypsin.
Max Ex/Em
Fluorophore Excitation Max (nm) Emission Max (nm)
FAM 494 520
Regulatory
For research purposes only
Guarantee
Guaranteed until date of expiry. Please see product label.
Acknowledgements
FLISP is a trademark of Immunochemistry Technologies, LLC.

MULTIPLE STORAGE CONDITIONS APPLY ON ARRIVAL. Store the unopened kit (and each unopened component) according to the storage instructions on each component label.
Store the FLISP reagent at -20oC. Once reconstituted in DMSO, the FLISP stock should be stored at -20oC for 6 months, protected from light. Avoid repeated freezing and thawing.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Flow Cytometry
Immunofluorescence
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.

Instructions For Use
Instructions for use can be found at www.bio-rad-antibodies.com/uploads/IFU/APO009.pdf

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Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"