Recombinant
RabMAb

Recombinant Anti-Granzyme A antibody [EPR20161] (ab209205)

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Overview

  • Product name

    Anti-Granzyme A antibody [EPR20161]
    See all Granzyme A primary antibodies

  • Description

    Rabbit monoclonal [EPR20161] to Granzyme A

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human Granzyme A aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P12544

  • Positive control

    • WB: NK-92 whole cell lysate; Human spleen, tonsil, lymph node and lung lysates. IHC-P: Human NK/T lymphoma, spleen and endometrium 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 ab209205 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 1/1000. Detects a band of approximately 28 kDa (predicted molecular weight: 29 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/70.

Target

  • Function

    This enzyme is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Lys or Arg. Cleaves APEX1 after 'Lys-31' and destroys its oxidative repair activity. Involved in apoptosis.

  • Sequence similarities

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

  • Cellular localization

    Secreted. Cytoplasmic granule.
  • Information by UniProt

  • Database links

    • Alternative names

      • CTL tryptase antibody
      • CTLA3 antibody
      • Cytolytic t cell and natural killer cell specific trypsin like serine protease antibody
      • Cytotoxic T lymphocyte associated serine esterase 3 antibody
      • Cytotoxic T lymphocyte proteinase 1 antibody
      • Cytotoxic T-lymphocyte proteinase 1 antibody
      • Fragmentin 1 antibody
      • Fragmentin-1 antibody
      • GRAA_HUMAN antibody
      • Granzyme 1 cytotoxic T lymphocyte associated serine esterase 3 antibody
      • Granzyme A (Cytotoxic T lymphocyte associated serine esterase 3; Hanukah factor serine protease) antibody
      • Granzyme A (granzyme 1, cytotoxic T lymphocyte associated serine esterase 3) antibody
      • Granzyme A antibody
      • Granzyme A precursor antibody
      • Granzyme-1 antibody
      • GZMA antibody
      • H factor antibody
      • Hanukah factor serine protease antibody
      • Hanukkah factor antibody
      • HF antibody
      • HFSP antibody
      • TSP1 antibody
      see all

    Images

    • Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)
      All lanes : Anti-Granzyme A antibody [EPR20161] (ab209205) at 1/10000 dilution

      Lane 1 : NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) whole cell lysate
      Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 29 kDa
      Observed band size: 28 kDa
      why is the actual band size different from the predicted?


      Exposure time: 1 second


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression profile is consistent with what has been described in the literature (PMID:15911377).

    • Immunocytochemistry/ Immunofluorescence - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Immunocytochemistry/ Immunofluorescence - Anti-Granzyme A antibody [EPR20161] (ab209205)

      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 A with ab209205 at 1/100 dilution, followed by Goat anti-rabbit IgG (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 ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

      Immunohistochemical analysis of paraffin-embedded human NK/T lymphoma tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Cytoplasmic staining on human NK/T lymphoma 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 A antibody [EPR20161] (ab209205)
      Flow Cytometry - Anti-Granzyme A antibody [EPR20161] (ab209205)

      Flow cytometric analysis of 4% paraformaldehyde-fixed NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells labeling Granzyme A with ab209205 at 1/70 dilution (red) compared with a rabbit monoclonal IgG 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.

    • Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)
      All lanes : Anti-Granzyme A antibody [EPR20161] (ab209205) at 1/1000 dilution

      Lane 1 : Human spleen lysate at 10 µg
      Lane 2 : Human tonsil lysate at 10 µg
      Lane 3 : Human lymph node lysate at 10 µg
      Lane 4 : Human lung lysate at 20 µg

      Secondary
      Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
      Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 29 kDa
      Observed band size: 28 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 15 seconds, Lane 2: 3 minutes, Lane 3: 10 seconds, Lane 4: 5 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

      Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Cytoplasmic staining on human spleen is observed [PMID: 9637500].

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

      Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Cytoplasmic staining on some stromal cells of human endometrium 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.

    References

    This product has been referenced in:

    • Wakiyama H  et al. Cytolytic Activity (CYT) Score Is a Prognostic Biomarker Reflecting Host Immune Status in Hepatocellular Carcinoma (HCC). Anticancer Res 38:6631-6638 (2018). Read more (PubMed: 30504371) »
    See 1 Publication for this product

    Publishing research using ab209205? Please let us know so that we can cite the reference in this datasheet.

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