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  1. Link

    granzyme-b-antibody-epr22645-206-ab255598.pdf

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Cell Biology Apoptosis Extracellular Signals Granzymes
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RecombinantRabMAb

Recombinant Anti-Granzyme B antibody [EPR22645-206] (ab255598)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (3)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Immunoprecipitation - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Western blot - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
  • Anti-Granzyme B antibody [EPR22645-206] (ab255598)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22645-206] to Granzyme B
  • Suitable for: WB, IHC-P, IHC-Fr, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Granzyme B antibody [EPR22645-206]
    See all Granzyme B primary antibodies
  • Description

    Rabbit monoclonal [EPR22645-206] to Granzyme B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse splenocyte lysate. IHC-P: Human tonsil and colon tissues; Mouse lung and spleen tissue. IHC-Fr: Mouse spleen tissue; Rat spleen tissue. IP: Mouse spleen 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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22645-206
  • 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 [EPR22645-206] - BSA and Azide free (ab255868)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Recombinant Protein

    • Recombinant Human Granzyme B protein (ab158621)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab255598 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 27, 65 kDa (predicted molecular weight: 27 kDa).
IHC-P (3)
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/200.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 27, 65 kDa (predicted molecular weight: 27 kDa).
IHC-P
1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/200.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

IP
1/30.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

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
    • Entrez Gene: 14939 Mouse
    • Entrez Gene: 171528 Rat
    • Omim: 123910 Human
    • SwissProt: P10144 Human
    • SwissProt: P04187 Mouse
    • SwissProt: P18291 Rat
    • Unigene: 1051 Human
    • Unigene: 132115 Rat
    • Unigene: 145789 Rat
    • Unigene: 21395 Rat
    see all
  • 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 [EPR22645-206] (ab255598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in human tonsil (PMID: 29042640) is observed.

    The section was incubated with ab255598 for 30 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in human colon is observed.

    The section was incubated with ab255598 for 30 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling ab255598 with Granzyme B at 1/200 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution. The nuclear counterstain was DAPI (Blue).

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat spleen tissue labeling ab255598 with Granzyme B at 1/200 dilution followed by ab150077 AlexaFluor® 488 Goat anti-Rabbit secondary at 1/1000 dilution. The nuclear counterstain was DAPI (Blue).

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor 488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Granzyme B with ab255598 at 1/3000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in mouse lung (PMID: 23642129) is observed.

    The section was incubated with ab255598 for 30 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Granzyme B with ab255598 at 1/3000 dilution  followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on lymphocytes in mouse spleen (PMID: 7697916) is observed.

    The section was incubated with ab255598 for 30 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    Counterstained with Hematoxylin.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

  • Immunoprecipitation - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Immunoprecipitation - Anti-Granzyme B antibody [EPR22645-206] (ab255598)

    Granzyme B was immunoprecipitated from 0.35 mg mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate with ab255598 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab255598 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

    Lane 1: Mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate 10μg

    Lane 2: ab255598 IP in mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255598 in mouse spleen cells (treated with 2.5ug/ml Concanavalin A for 72h) whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

  • Western blot - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Western blot - Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    All lanes : Anti-Granzyme B antibody [EPR22645-206] (ab255598) at 1/1000 dilution

    Lane 1 : Untreated mouse splenocyte lysate
    Lane 2 : Mouse splenocytes treated with 2.5µg/ml concanavalin for 72 hours, lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 27 kDa
    Observed band size: 28,65 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    The expression of Granzyme B is elevated in stimulated splenocytes. Granzyme B forms complex with its protease inhibitor.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 8663264, 17116752, 21919028, 9774654).

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

  • Anti-Granzyme B antibody [EPR22645-206] (ab255598)
    Anti-Granzyme B antibody [EPR22645-206] (ab255598)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

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 (3)

Publishing research using ab255598? Please let us know so that we can cite the reference in this datasheet.

ab255598 has been referenced in 3 publications.

  • Scopim-Ribeiro R  et al. NSG Mice Facilitate ex vivo Characterization of Ewing Sarcoma Lung Metastasis Using the PuMA Model. Front Oncol 11:645757 (2021). PubMed: 33828989
  • Wu SY  et al. MYC suppresses STING-dependent innate immunity by transcriptionally upregulating DNMT1 in triple-negative breast cancer. J Immunother Cancer 9:N/A (2021). PubMed: 34321275
  • Luo F  et al. Lymphocyte activating gene 3 protein expression in nasopharyngeal carcinoma is correlated with programmed cell death-1 and programmed cell death ligand-1, tumor-infiltrating lymphocytes. Cancer Cell Int 21:458 (2021). PubMed: 34454491

Customer reviews and Q&As

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1-3 of 3 Abreviews

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Granzyme B antibody [EPR22645-206]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (palatine tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Permeabilization
No
Specification
palatine tonsil
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Hannes Kaddatz

Verified customer

Submitted Mar 27 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Granzyme B antibody [EPR22645-206]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Hannes Kaddatz

Verified customer

Submitted Dec 31 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Granzyme B antibody [EPR22645-206]

Average
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (COLON)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA
Permeabilization
No
Specification
COLON
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 28 2022

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