Human Granzyme A Antibody

Catalog #: AF2905 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF2905
AF2905-SP

Detection of Human Granzyme A by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Granzyme A Antibody Summary

Species Reactivity

Human

Specificity

Detects human Granzyme A in direct ELISAs.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Granzyme A
Cys26-Val262
Accession # P12544

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

0.25 µg/mL

See below

ELISA

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.

Immunocytochemistry

5-15 µg/mL

See below

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

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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.

Immunocytochemistry

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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.

ELISA

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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).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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 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.

References

Lieberman, J. and Z. Fan (2003) Curr. Opin. Immunol. 15:553.
Gershenfeld, H.K. et al. (1988) Proc. Natl. Acad. Sci. USA 85:1184.

Entrez Gene IDs

3001 (Human); 14938 (Mouse); 266708 (Rat)

Alternate Names

CTL Tryptase; CTLA3; CTLA3Granzyme A (Cytotoxic T-lymphocyte-associated serine esterase-3; Hanukah factorserine protease); Cytotoxic T-lymphocyte proteinase 1; EC 3.4.21; Fragmentin-1; granzyme A (granzyme 1, cytotoxic T-lymphocyte-associated serine esterase 3); Granzyme A; Granzyme-1; GZMA; H factor; Hanukkah factor; HF; HFSP; HFSPEC 3.4.21.78