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Granzyme a

Manufactured by R&D Systems
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Granzyme A is a serine protease enzyme found in the cytotoxic granules of natural killer cells and cytotoxic T lymphocytes. It plays a key role in the induction of apoptosis in target cells during the immune response.

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Lab products found in correlation

Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Granulysin, by R&D Systems (1 mentions) Wizard 2480, by PerkinElmer (1 mentions) Cathepsin b, by R&D Systems (1 mentions) Granzyme b, by R&D Systems (1 mentions)

3 protocols using granzyme a

1

Quantifying Neuroinflammatory Biomarkers

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The following proteins were quantified using enzyme-linked immunosorbent assay kits: neurofilament light chain (NfL) (Human Diagnostics, Umea, Sweden); CXCL13/BLC/BCA-1, granzyme A, and granulysin (R&D System, MN); chitinase 3 like 1 (MicroVue, Athens, OH); perforin, granzyme B, and granzyme H (Invitrogen–Thermo Fisher Scientific, MA), were used according to the manufacturer's instructions.
Cruciani C., Puthenparampil M., Tomas-Ojer P., Jelcic I., Docampo M.J., Planas R., Manogaran P., Opfer R., Wicki C., Reindl M., Jelcic I., Lutterotti A., Martin R, & Sospedra M. (2021). T-Cell Specificity Influences Disease Heterogeneity in Multiple Sclerosis. Neurology® Neuroimmunology & Neuroinflammation, 8(6), e1075.
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2

Granzyme B Activation and Inhibition Assay

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Pro-granzyme B (R&D Systems) was activated to granzyme B using cathepsin B (R&D systems) following the manufacturer’s protocol. Enzyme activity was assessed by cleavage of BOC-Ala-Ala-Asp-SBZL (Sigma) and reaction with DTNB (Sigma) with absorbance measurements at 405 nm (NanoDrop 2000 Spectrophotometer, Thermo Scientific). Serial dilutions of non-radioactive gallium-labeled NOTA-GZP were used to inhibit granzyme B by incubation at 37°C for 30 min prior to addition of substrate. Additionally, 68Ga-NOTA-GZP was incubated with the activated forms of granzyme B, granzyme A (R&D Systems), granzyme H (R&D Systems), granzyme K (Enzo Life Sciences) and pro-granzyme B to assess the specificity of the peptide. After a 30 min incubation at 37°C, enzymes were purified by size exclusion chromatography and bound radioactivity assessed by gamma counter (Wizard 2480, Perkin Elmer).
Larimer B.M., Wehrenberg-Klee E., Dubois F., Mehta A., Kalomeris T., Flaherty K., Boland G, & Mahmood U. (2017). Granzyme B PET Imaging as a Predictive Biomarker of Immunotherapy Response. Cancer research, 77(9), 2318-2327.
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3

Cytokine and Cytotoxic Protein Analysis

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The supernatants from the cell cultures were collected, centrifuged and stored at -80 °C until used for enzyme-linked immunosorbent assay (ELISA). The protein levels of interferon-γ (IFNγ), tumor necrosis factor-α (TNFα), granzyme A, granzyme B (R&D Systems, Minneapolis, MN), and perforin (Abcam, France) in the supernatant were determined using ELISA techniques according to the manufacturer’s instruction.
Yigitbilek F., Ozdogan E., Abrol N., Park W.D., Hansen M.J., Dasari S., Stegall M.D, & Taner T. (2022). Liver mesenchymal stem cells are superior inhibitors of NK cell functions through differences in their secretome compared to other mesenchymal stem cells. Frontiers in Immunology, 13, 952262.
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