Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)

Rabbit Recombinant Multiclonal Granzyme B antibody. Carrier free. Suitable for WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples.

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Images

Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (AB317459), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Multiclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	IP	ICC/IF	Flow Cyt (Intra)	IHC-Fr
Human	Tested	Tested	Tested	Tested	Tested	Not recommended
Mouse	Tested	Tested	Tested	Expected	Tested	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information GZMB

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

CGL1, CSPB, CTLA1, GRB, GZMB, Granzyme B, C11, CTLA-1, Cathepsin G-like 1, Cytotoxic T-lymphocyte proteinase 2, Fragmentin-2, Granzyme-2, Human lymphocyte protein, SECT, T-cell serine protease 1-3E, CTSGL1, Lymphocyte protease, HLP

Recommended products

Rabbit Recombinant Multiclonal Granzyme B antibody. Carrier free. Suitable for WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: RM1165
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab317459 is the carrier-free version of Anti-Granzyme B antibody [RM1165] ab317458.

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

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.

11 product images

Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse PBMC(mouse peripheral blood mononuclear cell) treated with 2.5ug/ml Concanavalin A for 72h (Bottom left and Bottom right) / Untreated mouse PBMC(Top left and Top right) cells labelling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/50 dilution (1 ug)/Top right and Bottom right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
mouse PBMC are co-stained with CD8a conjugated Pacific Blue (440/50BP).
Flow Cytometry (Intracellular) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC(human peripheral blood mononuclear cell) treated with 10ug/ml CD3 for 72hours and 10ug/ml CD28 for 72 hours(Bottom left and Bottom right) / Untreated Human PBMC(Top left and Top right) cells labelling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/50 dilution (1 ug)/Top right and Bottom right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
human PBMC are co-stained with CD8a conjugated Pacific Blue (440/50BP).
Immunoprecipitation - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Granzyme B was immunoprecipitated from 0.35 mg Mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate with Anti-Granzyme B antibody [RM1165] ab317458 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Granzyme B antibody [RM1165] ab317458 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate

Lane 2: Anti-Granzyme B antibody [RM1165] ab317458 IP in Mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Granzyme B antibody [RM1165] ab317458 in mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Granzyme B antibody [RM1165] (Anti-Granzyme B antibody [RM1165] ab317458) at 1/30 dilution
Lane 1: Mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate
Lane 2: Anti-Granzyme B antibody [RM1165] ab317458 IP in Mouse spleen cell treated with 2.5 ug/ml Concanavalin A for 72h whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 41s
Immunoprecipitation - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Granzyme B was immunoprecipitated from 0.35 mg KARPAS-299 (human T cell lymphoma cell) whole cell lysate with Anti-Granzyme B antibody [RM1165] ab317458 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Granzyme B antibody [RM1165] ab317458 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: KARPAS-299 (human T cell lymphoma cell) whole cell lysate

Lane 2: Anti-Granzyme B antibody [RM1165] ab317458 IP in KARPAS-299 (human T cell lymphoma cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Granzyme B antibody [RM1165] ab317458 in KARPAS-299 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Granzyme B antibody [RM1165] (Anti-Granzyme B antibody [RM1165] ab317458) at 1/30 dilution
Lane 1: KARPAS-299 (human T cell lymphoma cell) whole cell lysate
Lane 2: Anti-Granzyme B antibody [RM1165] ab317458 IP in KARPAS-299 (human T cell lymphoma cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Western blot - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Negative control: Jurkat (PMID:18437383).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-Granzyme B antibody [RM1165] (Anti-Granzyme B antibody [RM1165] ab317458) at 1/1000 dilution
Lane 1: KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg
Lane 2: SR (human pleural effusion lymphoblast) whole cell lysate at 20 µg
Lane 3: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 28 kDa, 15 kDa
Exposure time: 15s
Western blot - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
The molecular weight observed is consistent with what has been described in the literature (PMID: 8663264 and 17116752).
Granzyme B forms complex with its protease inhibitor (PMID:9774654).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-Granzyme B antibody [RM1165] (Anti-Granzyme B antibody [RM1165] ab317458) at 1/1000 dilution
Lane 1: Untreated mouse splenocyte lysate at 20 µg
Lane 2: Mouse splenocytes treated with 2.5µg/ml concanavalin for 72 hours lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 28 kDa, 36 kDa
Exposure time: 15s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on immune cells of mouse lung. The section was incubated with Anti-Granzyme B antibody [RM1165] ab317458 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse spleen. The section was incubated with Anti-Granzyme B antibody [RM1165] ab317458 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, 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 [RM1165] ab317458 at 1/1000 (0.51 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human tonsil. The section was incubated with Anti-Granzyme B antibody [RM1165] ab317458 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/1000 (0.51 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on immune cells of human colon. The section was incubated with Anti-Granzyme B antibody [RM1165] ab317458 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-Granzyme B antibody [RM1165] - BSA and Azide free (ab317459)
This data was developed using Anti-Granzyme B antibody [RM1165] ab317458, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized KARPAS-299 (human T cell lymphoma cell) cells labelling Granzyme B with Anti-Granzyme B antibody [RM1165] ab317458 at 1/50 (10.2 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in KARPAS-299 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Negative control: Jurkat (PMID:18437383).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.