Human Granzyme A Antibody Summary
Accession # P12544
This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human Granzyme A Monoclonal Antibody (Catalog # MAB29052).
This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Granzyme A DuoSet ELISA Kit (Catalog # DY2905-05) for convenient development of a sandwich ELISA.
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.
Granzyme A in Human PBMCs. Granzyme A was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human Granzyme A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2905) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Human Granzyme A ELISA Standard Curve. Recombinant Human Granzyme A protein was serially diluted 2-fold and captured by Mouse Anti-Human Granzyme A Monoclonal Antibody (Catalog # MAB29052) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Granzyme A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2905) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).
Detection of Human Granzyme A by Western Blot. Western blot shows lysates of NK-92 human natural killer lymphoma cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Granzyme A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2905) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Granzyme A at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
- 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 A
Granzyme A 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. Granzyme A is the most abundant protease in CTL and NK cells. It induces caspase‑independent cell death when introduced into target cells by perforin (1). Human granzyme A is synthesized as a 262 amino acid (aa) precursor with a signal peptide (aa 1‑26), a propeptide (aa 27‑28) and a mature chain (aa 29‑262) (2). After being activated by lysyl endopeptidase removal of the propeptide, Granzyme A cleaves thioester substrates. Mature human granzyme A shares 71% amino acid sequence identity with mouse granzyme A.
- Lieberman, J. and Z. Fan (2003) Curr. Opin. Immunol. 15:553.
- Gershenfeld, H.K. et al. (1988) Proc. Natl. Acad. Sci. USA 85:1184.
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