Human Granzyme A Antibody

Catalog # Availability Size / Price Qty
AF2905
AF2905-SP
Human Granzyme A Antibody in Western Blot
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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 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.

Data Examples

Western Blot Detection  of Human Granzyme A by Western Blot. View Larger

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 Granzyme A in Human PBMCs. View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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
  1. Lieberman, J. and Z. Fan (2003) Curr. Opin. Immunol. 15:553.
  2. 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

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Cell and Tissue Staining Kits

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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