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Rat Recombinant Monoclonal Granzyme B antibody. Carrier free. Suitable for WB, IHC-P, Flow Cyt (Intra) and reacts with Human samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (AB289895), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (AB289895), expandable thumbnail
  • Western blot - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (AB289895), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (AB289895), expandable thumbnail

Key facts

Isotype

IgG2a

Host species

Rat

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PFlow Cyt (Intra)
Human
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

Abundant protease in the cytosolic granules of cytotoxic T-cells and NK-cells which activates caspase-independent pyroptosis when delivered into the target cell through the immunological synapse (PubMed:1985927, PubMed:3262682, PubMed:3263427). It cleaves after Asp (PubMed:1985927, PubMed:8258716). Once delivered into the target cell, acts by catalyzing cleavage of gasdermin-E (GSDME), releasing the pore-forming moiety of GSDME, thereby triggering pyroptosis and target cell death (PubMed:31953257, PubMed:32188940). Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -9 and -10 (CASP3, CASP9 and CASP10, respectively) to give rise to active enzymes mediating apoptosis (PubMed:9852092). Cleaves and activates CASP7 in response to bacterial infection, promoting plasma membrane repair (By similarity).

Alternative names

Recommended products

Rat Recombinant Monoclonal Granzyme B antibody. Carrier free. Suitable for WB, IHC-P, Flow Cyt (Intra) and reacts with Human samples.

Key facts

Isotype

IgG2a

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR8260

Purification technique

Ion exchange chromatography

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab289895 is the carrier-free version of Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888.

This rat monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (Anti-Granzyme B antibody [EPR8260] ab134933). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Granzyme B also known as GZMB GB11 or granzyme B protein is a serine protease with a molecular mass of approximately 32 kDa. It is expressed mainly in cytotoxic T lymphocytes and natural killer (NK) cells. This enzyme plays a mechanical role in inducing apoptosis in target cells serving as an effector protein in the immune system's defense against virally infected cells or transformed cancer cells. The activity of granzyme B relies on its ability to cleave after aspartate residues in substrate proteins leading to the activation of apoptotic pathways.

Biological function summary

Granzyme B participates prominently in the immune response by activating caspases particularly caspase-3 which promotes the breakdown of cellular components necessary for apoptosis. Granzyme B does not function in isolation but acts in concert with other immune system factors such as perforin to effectively induce cell death. Perforin creates pores in the target cell membrane allowing granzyme B to enter and instigate the apoptosis sequence. The enzyme also contributes to the processing of cytokines which enhances the immune response further.

Pathways

Studies have determined that granzyme B is critical in the apoptosis pathway particularly in the granule exocytosis pathway. It closely interacts with proteins such as perforin and other granzymes to mediate apoptosis in target cells. Granzyme B also plays a role in the inflammatory response and can influence pathways associated with cytotoxic T cell signaling. Its pathway interactions ensure effective elimination of damaged or infected cells maintaining tissue homeostasis.

Associated diseases and disorders

Granzyme B has associations with autoimmune diseases and cancer. Abnormally high levels of granzyme B can contribute to tissue damage and inflammation in autoimmune conditions like rheumatoid arthritis. In the context of cancer granzyme B aids in tumor surveillance and destruction when functioning correctly but impaired granzyme B activity can lead to evasion of immune detection by cancerous cells. Perforin also plays a role in these conditions closely working with granzyme B to either protect against or drive disease progression.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895)

    This data was developed using Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Granzyme B with Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888 at 1/100 dilution, followed by ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human tonsil.

    The section was incubated with Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888 for 30 mins at room temperature and followed by rat IgG antibody (Rabbit Anti-Rat IgG H&L preadsorbed ab102248) for 8 mins.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection).

  • Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895)

    This data was developed using Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized No-GFP-CD16.NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells labelling Granzyme B with Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888 at 1/50 dilution (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rat IgG Fc (DyLight 488, ab96971) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895), expandable thumbnail

    Western blot - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895)

    This data was developed using Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888, the same antibody clone in a different buffer formulation.

    Negative control: Jurkat (PMID:18437383, 25168906

    5% NFDM/TBST was used as a blocking and diluting buffer.

    All lanes: Western blot - Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888) at 1/1000 dilution

    Lane 1: KARPAS-299 (human anaplastic large cell lymphoma), whole cell lysate at 20 µg

    Lane 2: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

    Lane 3: SR (human pleural effusion lymphoblast), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution

    Predicted band size: 28 kDa

    Observed band size: 28 kDa

    Exposure time: 48s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] - Chimeric – BSA and Azide free (ab289895)

    This data was developed using Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Granzyme B with Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888 at 1/100 dilution, followed by ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Negative control: no staining on the glomerulus and tubule epithelia in human kidney (PMID: 28900605).

    The section was incubated with Anti-Granzyme B antibody [EPR8260] - Rat IgG2a (Chimeric) ab289888 for 30 mins at room temperature and followed by rat IgG antibody (Rabbit Anti-Rat IgG H&L preadsorbed ab102248) for 8 mins.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins was used.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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