Overview

  • Product name

    Mouse Granzyme B ELISA Kit
    See all Granzyme B kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Mouse spleen 5 3.9%
    Inter-assay
    Sample n Mean SD CV%
    Mouse spleen 3 6.1%
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    24.4 pg/ml
  • Range

    188 pg/ml - 12000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98 94% - 101%
    Serum 94 89% - 103%
    Cell culture media 109 103% - 120%
    Citrate Plasma 90 85% - 96%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Granzyme B in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Granzyme B protein in mouse tissue supernatant, serum, and plasma (citrate) samples


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Granzyme B is an enzyme involved in activation of caspases during cell mediated immune responses. Granzyme B specifically cleaves proteins at the site after asparagine residues.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Granzyme B Capture Antibody 1 x 600µl
    10X Mouse Granzyme B Detector Antibody 1 x 600µl
    Mouse Granzyme B Lyophilized recombinant Protein 2 vials
    Antibody Diluent CPR 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    TMB Development Solution 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • 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
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab238265 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Granzyme B were measured in duplicates, interpolated from the Granzyme B standard curves and corrected for sample dilution. Undiluted samples are as follows: treated mouse spleen supernatant 1: 67 and untreated mouse spleen supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Granzyme B concentration was determined to be 340.6 ng/mL in treated mouse spleen supernatant and 31.7 ng/ml in untreated mouse spleen supernatant.

Protocols

References

ab238265 has not yet been referenced specifically in any publications.

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