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.

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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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR20129-217] (ab208586)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Immunocytochemistry/ Immunofluorescence - Anti-Granzyme B antibody [EPR20129-217] (ab208586)

      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.

    • Western blot - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Western blot - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      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.

    • Western blot - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Western blot - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      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
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR20129-217] (ab208586)

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

      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.

    Publishing research using ab208586? Please let us know so that we can cite the reference in this datasheet.

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