Detection of Human Hydroxyacid Oxidase‑1/HAO‑1 by Western Blot.
Western blot shows lysates of human liver tissue. PVDF Membrane was probed with 0.5 µg/mL of Sheep Anti-Human Hydroxyacid Oxidase‑1/HAO‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6197) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Hydroxyacid Oxidase‑1/HAO‑1 at approximately 41 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
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.
Background: Hydroxyacid Oxidase-1/HAO-1
Glycolate oxidase is a member of the superfamily of the alpha ‑hydroxy acid oxidases (HAO), enzymes that are present in both plants and animals (1). It catalyzes the FMN-mediated oxidation of glycolate to glyoxylate and glyoxylate to oxalate with reduction of oxygen to hydrogen peroxide (2, 3). The co‑factor, FMN, is tightly bound but not covalently linked to the protein. In humans and other vertebrates, HAOs are found primarily in the peroxisomes of liver, kidney, and pancreas. Three HAOs have been identified in humans (4). HAO-1 is most highly expressed in liver and pancreas and is most active on two-carbon substrates such as glycolate. HAO-2 is expressed in liver and kidney and has greater activity against long-chain alpha ‑hydroxy acid substrates such as 2‑hydroxypalmitate. HAO-3 is expressed primarily in the pancreas. Recently, HAO has been identified as a major contributor to hyperoxaluria, a disorder in which large deposits of calcium oxalate form kidney stones (5).
Caroline, V. et al. (2007) Arch. Biochem. Biophys. 465:410.
Murray, M.S. et al. (2008) Biochemistry, 47:2439.
Pennati, A. and G. Gadda. (2009) J. Biol. Chem. 284:31214.
Jones, J.M. et al. (2000) J. Biol. Chem. 275:12590.
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