15-PGDH/HPGD Protein, Human (C-His)

15-PGDH/HPGD, a short-chain nonmetalloenzyme alcohol dehydrogenase, is crucial for prostaglandin metabolism, influencing various physiological and cellular processes like inflammation. Mutations lead to hypertrophic osteoarthropathy. Biased expression in urinary bladder (RPKM 176.8) and stomach (RPKM 67.9), among other tissues. Multiple isoforms arise from various transcript variants. 15-PGDH/HPGD Protein, Human (C-His) is the recombinant human-derived 15-PGDH/HPGD, expressed by E. coli, with C-6*His labeled tag. The total length of 15-PGDH/HPGD Protein, Human (C-His) is 266 a.a..

15-PGDH/HPGD Protein, a member of the short-chain nonmetalloenzyme alcohol dehydrogenase protein family, plays a crucial role in the metabolism of prostaglandins, contributing to various physiological and cellular processes, including inflammation. Mutations in this gene are associated with primary autosomal recessive hypertrophic osteoarthropathy and cranioosteoarthropathy, underscoring its significance in maintaining normal skeletal development. The gene exhibits multiple transcript variants, encoding different isoforms, adding to the functional diversity of the encoded enzyme. Notably, 15-PGDH/HPGD demonstrates biased expression, with heightened levels observed in the urinary bladder (RPKM 176.8), stomach (RPKM 67.9), and 11 other tissues, suggesting its specialized roles in these tissues and its potential involvement in diverse physiological contexts.

Measure by the production of NADH during the oxidation of PGF2 alpha that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is 1500.536 pmol/min/μg.

Materials
Assay buffer: 50 mM Tris, 100 mM NaCl, 2 mM Dithiothreitol (DTT) , pH 9.0
15-PGDH/HPGD Protein, Human (C-His) (HY-P75547A)
Substrate: β-NAD, PGF2 α

Procedure
1. Prepare the substrate mixture:
a. Dilute β-NAD to 1 mM in assay buffer.
b. Dilute PGF2 α to 0.4 mM in assay buffer.
c. Mix equal volumes of each substance to a final concentration of 0.5 mM β-NAD and 0.2 mM PGF2 α.
2. Dilute Human HPGD to 1.0 ng/μL in assay buffer.
3. The reaction was started by adding 50 μL of 1.0 ng/μL Human HPGD and 50 μL of substrate mixture to each well, and 50 μL of assay buffer and 50 μL of substrate mixture to the control group.
4. The optical density (OD) was measured at 339 nm in kinetic mode for 5 minutes using a microplate reader.
5. Calculate Specific Activity：

Human

E. coli

C-6*His

NP_000851.2 (M1-Q266)

3248

Full Length

MHVNGKVALVTGAAQGIGRAFAEALLLKGAKVALVDWNLEAGVQCKAALDEQFEPQKTLFIQCDVADQQQLRDTFRKVVDHFGRLDILVNNAGVNNEKNWEKTLQINLVSVISGTYLGLDYMSKQNGGEGGIIINMSSLAGLMPVAQQPVYCASKHGIVGFTRSAALAANLMNSGVRLNAICPGFVNTAILESIEKEENMGQYIEYKDHIKDMIKYYGILDPPLIANGLITLIEDDALNGAIMKITTSKGIHFQDYDTTPFQAKTQ

Approximately 29 kDa

≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 10 % trehalose and 0.02% Tween 80.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.