Granzyme B Antibody (351927) [Unconjugated]

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 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.

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.

Granzyme B was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human Granzyme B Monoclonal Antibody (Catalog # MAB2906) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, CyTOF-ready, Flow, ICC/IF
• Clone: 351927
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Granzyme B Antibody (351927) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Granzyme B
  Gly19-Tyr247
  Accession # P10144
• Specificity: 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.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: GZMB
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Simple Western 5 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Granzyme B Antibody (351927) [Unconjugated]

• C11
• CCPI
• CGL1
• CGL-1
• CSPB
• CSP-B
• CTLA1
• CTLA-1
• CTSGL1
• Fragmentin-2
• Granzyme B
• Granzyme-2
• GranzymeB
• GRB
• GrzB
• GZMB
• HLP
• SECT

Background

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).

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Granzyme B Antibody (MAB2906)(4)

Publications using MAB2906

• Hélène Jamann, Qiao-Ling Cui, Haritha L. Desu, Florian Pernin, Olivier Tastet, Alexandre Halaweh et al. Contact-Dependent Granzyme B-Mediated Cytotoxicity of Th17-Polarized Cells Toward Human Oligodendrocytes Frontiers in Immunology 4/11/2022 [PMID: 35479072]
• Fadi Jacob, Ryan D. Salinas, Daniel Y. Zhang, Phuong T.T. Nguyen, Jordan G. Schnoll, Samuel Zheng Hao Wong et al. A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity Cell 1/1/2020 [PMID: 31883794]
• Ming‑Lei Zhang, Li Chen, Yan‑Jiao Li, Da‑Liang Kong PD‑L1/PD‑1 axis serves an important role in natural killer cell‑induced cytotoxicity in osteosarcoma Oncology Reports 9/3/2019 [PMID: 31485666]
• P Paidipally, D Tripathi, A Van, RK Radhakrish, R Dhiman, S Venkatasub, KP Devalraju, AR Tvinnereim, VL Valluri, R Vankayalap IL-21 regulates NK cell responses during Mycobacterium tuberculosis infection J. Infect. Dis., 2018-03-28;0(0):. 2018-03-28 [PMID: 29390153] (ICC, Human)

Bioinformatics

• Gene Symbol: GZMB
• Uniprot:
  • P10144()