15-PGDH/HPGD 蛋白, Mouse (His)

The 15-PGDH/HPGD protein catalyzes the NAD-dependent dehydrogenation of various hydroxylated polyunsaturated fatty acids (mainly eicosanoids and behenic acid). 15-PGDH/HPGD Protein, Mouse (His) is the recombinant mouse-derived 15-PGDH/HPGD protein, expressed by E. coli , with C-His labeled tag.

15-PGDH/HPGD 蛋白在催化多种羟基化多不饱和脂肪酸（主要是类二十烷酸和二十二烷酸，包括前列腺素、脂氧素和分解素）的 NAD 依赖性脱氢（氧化）过程中发挥着关键作用，从而形成其相应的酮（oxo）代谢物。这种酶活性对于调节细胞反应至关重要，特别是通过降低促增殖前列腺素（如前列腺素 E2）的水平。通过产生含氧脂肪酸产物，15-PGDH/HPGD 可以深刻影响细胞功能并抵消某些细胞因子的炎症作用。此外，该酶在灭活分解素方面发挥着重要作用，包括分解素 E1、D1 和 D2，这些分解素参与急性炎症和肥胖引起的脂肪炎症的消退阶段。

Measure by the production of NADH during the oxidation of PGF2 alpha of 0.4 mM that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is 1511.254 pmol/min/μg.

Materials
Assay buffer: 50 mM Tris, 100 mM NaCl, 2 mM Dithiothreitol (DTT) , pH 9.0
15-PGDH/HPGD Protein, Mouse (His) (HY-P75555)
Substrates: beta-NAD, PGF2 alpha

Procedure
1. Prepare the substrate mixture:
a. Dilute beta-NAD to 1 mM in the assay buffer.
b. Dilute PGF2 alpha to 0.4 mM in the assay buffer.
c. Mix equal volumes of each solution to achieve a final concentration of 0.5 mM beta-NAD and 0.2 mM PGF2 alpha.
2. Dilute HPGD to 10 ng/μL in the assay buffer.
3. Add 50 μL of 1.0 ng/μL HPGD and 50 μL of the substrate mixture to each well to start the reaction. For the control group, add 50 μL of assay buffer and 50 μL of the substrate mixture.
4. Using a microplate reader, measure in kinetic mode at 339 nm for 5 minutes.
5. Calculate the specific activity:

Mouse

E. coli

C-6*His

Q8VCC1 (M1-S269)

15446  [NCBI]

Full Length

MHVNGKVALVTGAAQGIGKAFAEALLLHGAKVALVDWNLEAGVKCKAALDEQFEPQKTLFVQCDVADQKQLRDTFRKVVDHFGRLDILVNNAGVNNEKNWEQTLQINLVSVISGTYLGLDYMSKQNGGEGGIIINMSSLAGLMPVAQQPVYCASKHGIIGFTRSAAMAANLMKSGVRLNVICPGFVDTPILESIEKEENMGQYIEYKDQIKAMMKFYGVLHPSTIANGLINLIEDDALNGAIMKITASKGIHFQDYDISPLLVKAPLTS

Approximately 30 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

无菌粉末.

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose, 8% mannitol 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.