|Expression cDNA Clone and Sequence
Recombinant protein was produced with TrueORF clone, RC216061. Click on the TrueORF clone link to view cDNA and protein sequences.|
||Predicted MW:||55.5 kDa|
|Purity:||> 80% as determined by SDS-PAGE and Coomassie blue staining|
|Concentration:||>50 ug/mL as determined by microplate BCA method|
|Buffer and Storage:||25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol.|
||Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps.
||The enzymatic activity of TP316061 (GBA) was measured by its ability to hydrolyze a fluorescent substrate 4-methylumbelliferyl-ß-D-glucopyranoside. The specific activity is > 70,000 pmol/hour/µg, as measured under the following conditions: 27 ng of GBA was incubated with 10 mM 4-methylumbelliferyl- ß-D-glucopyranoside in the following buffer at 37°C for 40 min: 150 mM citrate-phosphate buffer, pH 5.4, 0.25% (w/w) sodium taurocholate, 0.25% (w/w) Triton X-100, and 1% bovine serum albumin. The reaction was terminated by adding 0.5 volume of 1M glycine buffer, pH 12.5. The hydrolyzed product of reaction, 4-methylumbelliferone (4-MU), was measured using a FlexStation 3 microplate reader (Ex365/Em445). Specific activity of GBA was calculated based on a standard curve of known concentration of 4-MU.
||Other glycan degradationSphingolipid metabolismMetabolic pathwaysLysosome