Granzyme B Rabbit Polyclonal Antibody

• Catalog Number: orb10738
• Category: Antibodies
• Description: Granzyme B Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Mouse, Porcine, Rat
• Reactivity: Human
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GZMB (21-130/247aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 25 kDa
• Target: GZMB
• UniProt ID: P10144
• RRID: AB_10752790
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: GranzymeB; Granzyme-B; C11; Cathepsin G-like 1; CT
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Reeves, Kirsten M. et al. 18F-FMISO PET Imaging Identifies Hypoxia and Immunosuppressive Tumor Microenvironments and Guides Targeted Evofosfamide Therapy in Tumors Refractory to PD-1 and CTLA-4 Inhibition Clin Cancer Res, 28, 327-337 (2022)

PubMed: 34615724

Blank control: U937. Primary Antibody (green line): Rabbit Anti-Granzyme B antibody (orb10738), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at PBST. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Sample: Lane 1: Human K562 cell lysates, Lane 2: Human MOLT4 cell lysates, Primary: Anti-Granzyme B (orb10738) at 1/500 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 25 kDa, Observed band size: 35 kDa.

Tissue/Cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Granzyme B Polyclonal Antibody, Unconjugated (orb10738) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.