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-Leu-CMK Serine Protease Assay Kit
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  • Product Type
    Kits
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    APO007F, IFdatasheet pdfdatasheet pdf datasheet pdf datasheet pdf datasheet pdf datasheet pdf25 Tests
    APO007
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
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    • 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.
    • Intended Use
    • Product Form
    • Reconstitution
    • Preparation
    • Preservative Stabilisers
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
      FAM-Leu-CMK FLISP Reagent - lyophilized
      10X Wash Buffer, 15 mL
      Fixative, 6 mL
      Propidium Iodide, 1 mL
      Hoechst Stain, 1 mL
    • Preparing The Antibody
    • Test Principle
      Active chymotrypsin-like enzymes within the cell bind covalently to the FLISP FAM-Leu-CMK 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-Leu-CMK probe inhibits chymotrypsin C.
    • Buffer Solution
    • Storage
      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.
    • Shelf Life
      Please see label for expiry date
    • Acknowledgements
      FLISP is a trademark of Immunochemistry Technologies, LLC.
    • Regulatory
      For research purposes only
    • 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 NameYesNoMin DilutionMax 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.

    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use
      Instructions for use can be found at www.bio-rad-antibodies.com/uploads/IFU/APO007.pdf

    Additional Serine Protease Assay Kit Formats

    Formats Applications Sizes available
    Serine Protease Assay Kit : FAM (Green Fluorescence) F, IF 25 Tests
    • Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          FLICA™ 660 Poly Caspase KitICT912025 - 50 TestsF, IF
          ICT9120
          FAM FLICA™ Poly Caspase KitICT092100 TestsF, IF
          ICT092
          FAM FLICA™ Poly Caspase KitICT09125 TestsF, IF
          ICT091
          SR FLICA™ Poly Caspase KitICT917100 TestsF, IF
          ICT917
          SR FLICA™ Poly Caspase KitICT91625 TestsF, IF
          ICT916

          Recommended Positive Controls

            Histology Controls

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