HSD17B4 Protein, Human (P.pastoris, His)

HSD17B4 Protein, Human (P.pastoris, His) is the recombinant human-derived HSD17B4, expressed by P. pastoris , with N-6*His labeled tag. ,

HSD17B4 is a bifunctional enzyme involved in the peroxisomal fatty acid beta-oxidation pathway. It catalyzes two key reactions in fatty acid degradation: the hydration of 2-enoyl-CoA (trans-2-enoyl-CoA) to produce (3R)-3-hydroxyacyl-CoA, and the dehydrogenation of (3R)-3-hydroxyacyl-CoA to yield 3-ketoacyl-CoA (3-oxoacyl-CoA), which is further metabolized by SCPx. This enzyme can utilize straight-chain and branched-chain fatty acids, as well as bile acid intermediates as substrates. by similar: HSD17B4 plays a critical role in fatty acid metabolism, particularly within peroxisomes.

Measured by dehydrogenase activity through NADH production. The specific activity is 89.462 pmol/min/μg.

Materials
Detection Buffer: 50 mM Tris-HCl, 100 mM NaCl, pH 8.5, 200 μM NAD⁺
Substrate: 20 mM Estradiol (HY-B0141)
HSD17B4 Protein, Human (P.pastoris, His) (HY-P702863)
96-well Transparent Plate

Procedure
1. Dilute NADH standard with ddH₂O to 0, 0.03125, 0.0625, 0.125, 0.25, 0.5, 1 mmol/L. Add 100 μL of standard to each microplate well. Measure the OD value at 340 nm in endpoint mode. Plot a standard curve with the OD value as the ordinate and the amount of standard in pmol as the abscissa, and obtain the standard curve formula.
2. Reconstitute Human HSD17B4 protein to 50 μg/mL.
3. Prepare the protein system mixture by mixing 25 μL of 50 μg/mL Human HSD17B4 protein with 25 μL of detection buffer.
4. Dilute 20 mM estradiol to 40 μM using detection buffer.
5. Experimental group: 50 µL of 40 μM estradiol + 50 µL of protein system mixture; Control group: 50 µL of 40 μM estradiol + 50 µL of detection buffer.
6. Zero the microplate reader using the OD value of the blank group, and read the absorbance at 340 nm for 10 min in kinetic mode.
7. Calculate the specific activity:

Human

P. pastoris

N-6*His

P51659 (M1-L736)

3295

Full Length

MGSPLRFDGRVVLVTGAGAGLGRAYALAFAERGALVVVNDLGGDFKGVGKGSLAADKVVEEIRRRGGKAVANYDSVEEGEKVVKTALDAFGRIDVVVNNAGILRDRSFARISDEDWDIIHRVHLRGSFQVTRAAWEHMKKQKYGRIIMTSSASGIYGNFGQANYSAAKLGLLGLANSLAIEGRKSNIHCNTIAPNAGSRMTQTVMPEDLVEALKPEYVAPLVLWLCHESCEENGGLFEVGAGWIGKLRWERTLGAIVRQKNHPMTPEAVKANWKKICDFENASKPQSIQESTGSIIEVLSKIDSEGGVSANHTSRATSTATSGFAGAIGQKLPPFSYAYTELEAIMYALGVGASIKDPKDLKFIYEGSSDFSCLPTFGVIIGQKSMMGGGLAEIPGLSINFAKVLHGEQYLELYKPLPRAGKLKCEAVVADVLDKGSGVVIIMDVYSYSEKELICHNQFSLFLVGSGGFGGKRTSDKVKVAVAIPNRPPDAVLTDTTSLNQAALYRLSGDWNPLHIDPNFASLAGFDKPILHGLCTFGFSARRVLQQFADNDVSRFKAIKARFAKPVYPGQTLQTEMWKEGNRIHFQTKVQETGDIVISNAYVDLAPTSGTSAKTPSEGGKLQSTFVFEEIGRRLKDIGPEVVKKVNAVFEWHITKGGNIGAKWTIDLKSGSGKVYQGPAKGAADTTIILSDEDFMEVVLGKLDPQKAFFSGRLKARGNIMLSQKLQMILKDYAKL

86 kDa

Greater than 90 % as determined by reducing SDS-PAGE.

Lyophilized powder

Lyophilized from a 0.22 μm filtered solution of PBS, 6% Trehalose, pH 7.4.

<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.