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Granzyme B Activity Assay Kit (Fluorometric) (ab157403)

Submit a review Q&A (1)
AFC Standard Curve

    Key features and details

    • Assay type: Enzyme activity (quantitative)
    • Detection method: Fluorescent
    • Platform: Microplate reader
    • Sample type: Cell Lysate

    Overview

    • Product name

      Granzyme B Activity Assay Kit (Fluorometric)
      See all Granzyme B kits

    • Detection method

      Fluorescent

    • Sample type

      Cell Lysate

    • Assay type

      Enzyme activity (quantitative)

    • Species reactivity

      Reacts with: Mammals, Other species

    • Product overview

      In Abcam's Granzyme B Activity Assay Kit (Fluorometric), active human Granzyme B enzyme hydrolyzes the specific substrate to release the quench of fluorescent group, which can be detected fluorometrically at Ex/Em = 380/500 nm. This Assay Kit is rapid, reliable and sensitive and can detect Granzyme B activity in various biological samples.

    • Notes

      Granzyme B (GZMB, EC number 3.4.21.79) also known as Granzyme-2 is a serine protease stored in the granules of activated cytotoxic T cells and NK cells. Upon target cell contact, Granzyme B is directionally exocytosed and with the assistance of perforin enters the target cell. With its unique substrate specificity (Granzyme B prefers an aspartic acid residue at the P1 site of its substrates), Granzyme B processes and activates various pro-caspases thereby inducing apoptosis in the target cell.

       

    • Platform

      Microplate reader

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components Identifier 100 tests
      AFC Standard Yellow 1 x 0.1ml
      Granzyme B Assay Buffer WM 1 x 25ml
      Granzyme B Positive Control Green 1 vial
      Granzyme B Substrate Red 1 x 0.5ml

    • Research areas

    • Function

      This enzyme is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Asp. Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -7, -9 and 10 to give rise to active enzymes mediating apoptosis.

    • Sequence similarities

      Belongs to the peptidase S1 family. Granzyme subfamily.
      Contains 1 peptidase S1 domain.

    • Cellular localization

      Cytoplasmic granule. Cytoplasmic granules of cytolytic T-lymphocytes and natural killer cells.
    • Information by UniProt

    • Alternative names

      • C11
      • Cathepsin G like 1
      • Cathepsin G-like 1
      • CCPI
      • CGL 1
      • CGL1
      • CSP B
      • CSPB
      • CTLA 1
      • CTLA-1
      • CTLA1
      • CTSGL1
      • Cytotoxic serine protease B
      • Cytotoxic T lymphocyte associated serine esterase 1
      • Cytotoxic T lymphocyte proteinase 2
      • Cytotoxic T-lymphocyte proteinase 2
      • Fragmentin 2
      • Fragmentin-2
      • GRAB_HUMAN
      • Granzyme 2
      • Granzyme B
      • Granzyme B (granzyme 2, cytotoxic T lymphocyte associated serine esterase 1)
      • Granzyme-2
      • GranzymeB
      • GRB
      • Gzmb
      • Hlp
      • Human lymphocyte protein
      • Lymphocyte protease
      • Protease, serine, B
      • SECT
      • T cell serine protease 1 3E
      • T cell serine protease 1-3E
      • T-cell serine protease 1-3E
      see all

    Images

    • AFC Standard Curve
      AFC Standard Curve

    References (0)

    Publishing research using ab157403? Please let us know so that we can cite the reference in this datasheet.

    ab157403 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Question

    First what is the substrate? I ask because often IEPD or IETD are used as substrates for Granzyme B, but these are also cleaved by caspases. Do you have a more complete protocol? Specifically for sample preparation what lysis buffers/conditions do you recommend?

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    Answer

    Due to proprietary restrictions I cannot disclose what exactly the substrate is in this kit, but I can assure you that it is granzyme B specific.


    Here is the protocol for processing cells for this assay:

    For cell samples: Start with ˜2X106 cells, suspend the cell pellet 500 μl (or ˜4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the supernatant for your subsequent assays. Appropriate dilutions of the sample must be tested in order to ensure the readings will fall within the linear range of the standard curve.

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