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  1. Link

    granzyme-b-antibody-epr20129-217-ab208586.pdf

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Cell Biology Apoptosis Extracellular Signals Granzymes
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RecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
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Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Western blot - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
  • 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)
  • Anti-Granzyme B antibody [EPR20129-217] (ab208586)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20129-217] to Granzyme B
  • Suitable for: WB, IHC-P, mIHC
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free PE

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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, IHC-P, mIHCmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human Granzyme B recombinant protein; IHC-P: Human colon and cervix cancer tissues. mIHC: Human breast cancer tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20129-217
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Extracellular Signals
    • Granzymes
    • Immunology
    • Adaptive Immunity
    • T Cells
    • Cytotoxic Cells
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Immunology
    • Innate Immunity
    • NK Cells
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Granzyme B antibody [EPR20129-217] - BSA and Azide free (ab219803)
    • PE Anti-Granzyme B antibody [EPR20129-217] (ab225471)
    • Alexa Fluor® 488 Anti-Granzyme B antibody [EPR20129-217] (ab225472)
    • Alexa Fluor® 647 Anti-Granzyme B antibody [EPR20129-217] (ab225473)
  • Assay kits

    • Granzyme B Activity Assay Kit (Fluorometric) (ab157403)
    • Granzyme B Inhibitor Screening Assay Kit (ab157405)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant mouse Granzyme B protein (ab50114)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

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).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
mIHC
Use at an assay dependent concentration.
Notes
WB
1/10000. Detects a band of approximately 28 kDa (predicted molecular weight: 28 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
mIHC
Use at an assay dependent concentration.
Application notes
Is unsuitable for Flow Cyt.

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.
  • Target information above from: UniProt accession P10144 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3002 Human
    • Omim: 123910 Human
    • SwissProt: P10144 Human
    • Unigene: 1051 Human
    • 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

    • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • 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.

    • 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.

    • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)
      Multiplex immunohistochemistry - Anti-Granzyme B antibody [EPR20129-217] (ab208586)This image is courtesy of ImmunoAtlas.

      Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

      Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

      The immunostaining was performed on a Leica Biosystems BOND® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences®).

      The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
      Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences®).

      This data is courtesy of ImmunoAtlas and it can be found here.

    • 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.

    • 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)

      Tissue Microarrays stained for "Anti-Granzyme B antibody [EPR20129-217]” using "ab208586" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab208586 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

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

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (2)

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

    ab208586 has been referenced in 2 publications.

    • Qiao T  et al. Inhibition of LDH-A by Oxamate Enhances the Efficacy of Anti-PD-1 Treatment in an NSCLC Humanized Mouse Model. Front Oncol 11:632364 (2021). PubMed: 33859941
    • Janiszewski T  et al. Noninvasive optical detection of granzyme B from natural killer cells with enzyme-activated fluorogenic probes. J Biol Chem 295:9567-9582 (2020). PubMed: 32439802

    Customer reviews and Q&As

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