Detection of Human Hexosaminidase A/ HEXA by Western Blot.
Western blot shows lysate of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Hexosaminidase A/HEXA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6237) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Hexosaminidase A/HEXA at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Hexosaminidase A/HEXA by Simple WesternTM.
Simple Western lane view shows lysate of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Hexosaminidase A/HEXA at approximately 59 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Hexosaminidase A/HEXA Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6237) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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: Hexosaminidase A/HEXA
HEXA (beta-hexosaminidase subunit alpha; also hexaminidase subunit alpha) is a 50-56 kDa glycoprotein member of the glycosyl hydrolase #20 family of enzymes. It is found in the lysosomes of a variety of cell types including fibroblasts, lacrimal acinar cells, and neurons. beta -Hexosaminidase is a multimeric enzyme that cleaves aminoacetylhexosamines from multiple glycosylated molecules. It exists in three noncovalent isozyme forms, among which is an A form that is trimeric and composed of one alpha - and two beta -subunits, a B form that is tetrameric and contains two alpha - and two beta -subunits, and a dimeric S form that contains two alpha -subunits. The alpha - and beta -subunits are products of related but distinct genes. The beta -hexosiminidase A form acts on a wide range of substrates, while the B form activity is biased towards sulfated hexoses. Human HEXA is synthesized as a 529 aa (amino acid) prepro-precursor. It contains a 22 aa signal sequence, a 66 aa prosegment, and a 441 aa mature region that contains a catalytic domain (aa 167-488). The prosegment may undergo additional processing through His90, and this segment (≈ 6 kDa) is known to remain linked to the mature region via a disulfide bond. Notably, circulating HEXA is comprised only of subunit proforms whose MW may run some 10 kDa higher in SDS-PAGE. Over aa 23-529, human HEXA shares 85% and 57% aa sequence identity with mouse HEXA and the human HEXA beta -subunit proprecursor, respectively.
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