Recombinant
RabMAb

Recombinant Anti-Granzyme B antibody [EPR20129-217] (ab208586)

Rabbit recombinant monoclonal Granzyme B antibody [EPR20129-217]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Human. Immunogen corresponding to recombinant full length protein.

Overview

  • Product name

    Anti-Granzyme B antibody [EPR20129-217]
    See all Granzyme B primary antibodies
  • Description

    Rabbit monoclonal [EPR20129-217] to Granzyme B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human Granzyme B aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P10144

  • Positive control

    • WB: Human Granzyme B recombinant protein; NK-92 whole cell lysate. IHC-P: Human colon and cervix cancer tissues. ICC/IF: NK-92 cells. Flow Cyt: NK-92 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208586 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
WB 1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 28 kDa).
ICC/IF 1/100.
Flow Cyt 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • 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
  • Database links

  • Alternative names

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

Images

  • Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling Granzyme B with ab208586 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on neutrophils and stroma cells of human cervix cancer is observed [PMID: 14512315]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NK-92 (Human peripheral blood malignant Non-Hodgkin's lymphoma cell line) cells labeling Granzyme B with ab208586 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NK-92 cell line. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-Granzyme B antibody [EPR20129-217] (ab208586) at 1/10000 dilution

    Lane 1 : Human Granzyme B recombinant protein
    Lane 2 : Human Granzyme H recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 28 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Granzyme B and human Granzyme H recombinant protein contain aa21-247 and aa21-246 with a His-tag. These two recombinant proteins were made in house.

  • Anti-Granzyme B antibody [EPR20129-217] (ab208586) at 1/10000 dilution + NK-92 (Human peripheral blood malignant Non-Hodgkin's lymphoma cell line) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 28 kDa
    Observed band size: 28 kDa


    Exposure time: 1 second
  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Granzyme B with ab208586 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on some stromal cells of human colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NK-92 (Human peripheral blood malignant Non-Hodgkin's lymphoma cell line) cells labeling Granzyme B with ab208586 at 1/500 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control(ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

References

ab208586 has not yet been referenced specifically in any publications.

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