A DNA construct of HexB-P2A-HexA-His tag is transfected into CHO cells. Processed HexA/B herterodimer is consisted of two subunits: human Hex B(Ala43-Met556) with C-terminus P2A tag, and human HexA (Leu23-Thr529) with C-terminus His tag. Heterodimer protein is purified by Nickel resin with gradient elution to separate HexA/B heterodimer from HexA homodimer.
Molecular Weight
Human HexA protein has the predicted molecular weight of 60 kD and human HexB protein has the predicted molecular weight of 60.6 kD . Under DTT-reducing conditions, both proteins migrate at approximately 65 kDa on SDS-PAGE.
Affinity Tag
C-His
Purity
>95% based on SDS-PAGE under reducing condition
Regulatory Status
RUO
Formulation
25 mM Tris, 150 mM NaCl, pH 7.5 (0.2 μm filtered)
Endotoxin level
Not tested
Protein Concentration
25µg size is bottled at 0.2mg/mL concentration. 100 µg size is supplied at a lot-specific concentration.
Storage and Handling
Briefly centrifuge the vial upon receipt. An unopened vial can be stored at 4°C for up to 2 weeks, or at -20°C or below for up to six months. The protein may be further diluted to 0.1 mg/mL using 0.22 µm-filtered 25 mM Tris, 150 mM NaCl, pH 7.5. For long-term storage, the diluted stock solution should be aliquoted and stored at ≤ –70°C to minimize freeze-thaw cycles. If additional dilution is required, carrier proteins such as FBS or BSA should be added to maintain protein stability.
Background Information
Beta-hexosaminidases are lysosomal enzymes that hydrolyze terminal N-acetyl-D-hexosamine residues from GM2 gangliosides and globo-sphingolipids. They exist in three isoforms: Hex A (αβ), Hex B (ββ), and Hex S (αα), formed by different combinations of α and β subunits encoded by the HEXA and HEXB genes. Recombinant HexB-P2A/HexA-His Heterodimer corresponds to Hex A isoform.
