Human Granzyme B Antibody

Catalog # Availability Size / Price Qty
MAB2906
MAB2906-SP

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Detection of Human Granzyme B by Western Blot.
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Citations (1)
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Human Granzyme B Antibody Summary

Species Reactivity
Human
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
Monoclonal Mouse IgG2A Clone # 351927
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Granzyme B
Gly19-Tyr247
Accession # P10144
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Simple Western
5 µg/mL
NK‑92 human natural killer lymphoma cell line
Immunohistochemistry
5-25 µg/mL
Immersion fixed paraffin-embedded sections of human tonsil
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
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
NK‑92 human natural killer lymphoma cell line fixed with paraformaldehyde and permeabilized with saponin

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Granzyme B antibody by Western Blot. View Larger

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.

Immunocytochemistry Granzyme B antibody in Human PBMCs by Immunocytochemistry (ICC). View Larger

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.

Simple Western View Larger

Detection of Human Granzyme B by Simple WesternTM. 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.

Immunohistochemistry View Larger

Detection of Granzyme B in human tonsil. 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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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
  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.
Entrez Gene IDs
3002 (Human); 14939 (Mouse)
Alternate Names
C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; CTLA1; CTLA-1; CTSGL1; Fragmentin-2; Granzyme B; Granzyme-2; GranzymeB; GRB; GrzB; GZMB; HLP; SECT

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Citation for Human Granzyme B Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. IL-21 regulates NK cell responses during Mycobacterium tuberculosis infection
    Authors: P Paidipally, D Tripathi, A Van, RK Radhakrish, R Dhiman, S Venkatasub, KP Devalraju, AR Tvinnereim, VL Valluri, R Vankayalap
    J. Infect. Dis., 2018-03-28;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC

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