Detection of Human GAPDH‑2 by Western Blot.
Western blot shows lysates of human testis tissue. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human GAPDH‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6276) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for GAPDH‑2 at approximately 56 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
GAPDH‑2 in Human Testes.
GAPDH‑2 was detected in immersion fixed paraffin-embedded sections of human testes using Goat Anti-Human GAPDH‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6276) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of sperm cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
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.
GAPDH-2 (Glyceraldehyde-3-phosphate dehydrogenase 2; also GAPD2 and GAPDS) is a 40-44 kDa member of the GAPDH family of enzymes. Due to a high incidence of proline residues, it may migrate anomalously at 56 kDa in SDS-PAGE. GAPDH-2 is expressed only in testis as a homotetramer, where it is found in the cytoplasm bound to the sperm fibrous sheath. Unlike GAPDH that has multiple functions, GAPDH-2 appears to participate only in carbohydrate metabolism, generating energy via glycolysis for sperm motility and fertility. Human GAPDH-2 is 408 amino acids (aa) in length. There are two NAD binding sites (Asp106 and Asn388) with a catalytic region between aa 223-226. One splice variant shows a nine aa insertion after Ser220. Over aa 75-222, human GAPDH-2 shares 84% aa identity with mouse GAPDH-2.
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