Detects human Granzyme B in direct ELISAs and Western blots. Does not cross-react with recombinant human (rh) Granzyme A, rhGranzyme H, recombinant mouse (rm) Granzyme B, rmGranzyme C, rmGranzyme D, or rmGranzyme G.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for Human Granzyme B Antibody

Detection of Human Granzyme B by Western Blot.

Western blot shows lysate of NK-92 human natural killer lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Granzyme B at approximately 32-35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Granzyme B in Human PBMCs.

Granzyme B was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human Granzyme B by Simple Western^TM.

Simple Western lane view shows lysates of NK-92 human natural killer lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Granzyme B at approximately 42 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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Applications for Human Granzyme B Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells
Sample: NK-92 human natural killer lymphoma cell line fixed with paraformaldehyde and permeabilized with saponin

Simple Western

5 µg/mL
Sample: NK‑92 human natural killer lymphoma cell line

Western Blot

0.5 µg/mL
Sample: NK‑92 human natural killer lymphoma cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Granzyme B

Granzyme B is a member of the granzyme family of the serine proteases found specifically in the cytotoxic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells (1, 2). Granzyme B plays an essential role in granule-mediated apoptosis and may have additional roles in rheumatoid arthritis and in bacterial and viral infections (3). It activates various caspases and cleaves proteins such as aggrecan (3). Human Granzyme B is synthesized as a precursor (247 residues) with a signal peptide (residues 1-18), a pro peptide (residues 19-20), and a mature chain (residues 21-247) (4-6). The recombinant human (rh) Granzyme B consisting of residues 19-247 was expressed and purified. After being activated by active cathepsin C, rhGranzyme B cleaves a thioester substrate described previously (3).

References

Kam, C-M. et al. (2000) Biochim. Biophys. Acta 1477:307.
Smyth, M.J. et al. (1996) J. Leukoc. Biol. 60:555.
Froelich, C.J. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. pp. 1549.
Schmid, J. and C. Weissman (1987) J. Immunol. 139:250.
Caputo, A. et al. (1988) J. Biol. Chem. 263:6363.
Trapani, J.A. et al. (1988) Proc. Natl. Acad. Sci. USA 85:6924.

Alternate Names

CGL-1, CGL1, CSPB, CTLA-1, CTLA1, CTSGL1, Fragmentin-2, Granzyme-2, GRB, GrzB, GZMB, HLP, SECT

Entrez Gene IDs

3002 (Human); 14939 (Mouse)

Gene Symbol

GZMB

UniProt

P10144

Additional Granzyme B Products

Product Documents for Human Granzyme B Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Granzyme B Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Granzyme B Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB2906

Key Product Details

Species Reactivity

Validated:

Cited:

Applications