Recombinant Anti-Granzyme B antibody [EPR22645-206] (ab255598)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-9-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-tonsil-human.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Recombinant Protein
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).
|
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| IHC-P | (2) |
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. |
Target
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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
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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
see all -
Alternative names
- C11 antibody
- Cathepsin G like 1 antibody
- Cathepsin G-like 1 antibody
see all
Images
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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).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-8-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-colon-human.jpg)
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)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-7-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-frozen-spleen-mouse.jpg)
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.
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![Immunohistochemistry (Frozen sections) - Anti-Granzyme B antibody [EPR22645-206] (ab255598)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-6-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-frozen-spleen-rat.jpg)
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)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-5-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-lung-mouse.jpg)
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)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-4-anti-granzyme-b-antibody-epr22645-206-immunohistochemistry-spleen-mouse.jpg)
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)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-3-anti-granzyme-b-antibody-epr22645-206-immunoprecipitation-spleen-mouse.jpg)
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
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![Western blot - Anti-Granzyme B antibody [EPR22645-206] (ab255598)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-2-anti-granzyme-b-antibody-epr22645-206-western-blot-splenocytes-mouse.jpg)
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 secondsThe 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.
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![Anti-Granzyme B antibody [EPR22645-206] (ab255598)](https://www.abcam.com/ps/products/255/ab255598/Images/ab255598-10-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
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