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Active Granzyme B (GZMB)

GZM-B; HLP; CTLA1; CCPI; CGL1; CSP-B; CSPB; CTSGL1; SECT; Granzyme 2; Cytotoxic T-Lymphocyte-Associated Serine Esterase 1; Fragmentin 2; Cytotoxic Serine Protease B

ACTIVITY TEST

Granzyme B (GZMB) is a serine protease predominantly secreted by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. It serves as a central effector molecule in the granule exocytosis pathway, which is essential for cell-mediated elimination of virus-infected and malignant cells. Upon target cell recognition, CTLs and NK cells release GZMB together with perforin. Perforin facilitates the entry of GZMB into the target cell by forming pores in the membrane, where GZMB then initiates apoptotic signaling. In addition to its well-established role in apoptosis, GZMB also contributes to immune regulation and tissue remodeling through involvement in inflammatory responses and extracellular matrix degradation. Besides,Caspase-3 has been identified as an interactor of GZMB, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant rat GZMB and recombinant rat Caspase-3 .Briefly, biotin-linked GZMB were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µl were then transferred to Caspase-3-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of GZMB and Caspase-3 was shown in Figure 1, the EC50 for this effect is 2.62µg/mL.

USAGE

Reconstitute in ddH2O to a concentration less than or equal to 0.1mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
International Journal of Cancer Effective immunotherapy of rat glioblastoma with prolonged intratumoral delivery of exogenous heat shock protein Hsp70 Pubmed:24691976
Biol Pharm Bull. Z-100, an Immunomodulatory Extract of Mycobacterium tuberculosis Strain Aoyama B, Prevents Spontaneous Lymphatic Metastasis of B16-BL6 Melanoma Pubmed:24818259
Journal of Dermatological Science Relationships among plasma granzyme B level, pruritus and dermatitis in patients with atopicdermatitis. pubmed:27686401
Oncotarget Tumor-derived granzyme B-expressing neutrophils acquire antitumor potential after lipid A treatment Pubmed:29983866
Theranostics A novel Granzyme B nanoparticle delivery system simulates immune cell functions for suppression of solid tumors Pubmed: 31695790
Am J Transl Res Combined treatment with epigenetic agents enhances anti-tumor activity of T cells by upregulating the ACRBP expression in hepatocellular carcinoma 34377237
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