ALDH1A1 蛋白, Human (His)

ALDH1A1 Protein, Human (His) is a human recombinant ALDH1A1 with an N-terminal Met and His tag produced in E. coli. ALDH1A1 Protein, Human (His) contains 501 amino acids (1-501 a.a.).

乙醛脱氢酶 1-A1 (ALDH1A1) 是乙醛脱氢酶家族的一部分。人 ALDH1A1 是一种胞质同源四聚体，在多种组织（皮肤、眼睛、肝脏、肾脏、睾丸和大脑）中表达。ALDH1A1 可氧化多种脂肪族和芳香族醛，包括内源性形成的醛类，例如视黄醛和 3,4-二羟基苯乙醛 (DOPAL)、异生物质（例如苯甲醛）或脂质过氧化过程中产生的醛类^[1]。

Measured by its ability to produce NADH during the oxidation of propionaldehyde. The specific activity is 77.63 pmol/min/µg, as measured under the described condition.

Materials
ALDH1A1 Protein, Human (His) (HY-P7474)
Detection Diluent: Ultra-pure water
1 M Tris Solution, pH 8.5
2 M KCl Solution
1 M DTT Solution
200 mM Propionaldehyde
100 mM β-NAD

Procedure
1. Dilute Human ALDH1A1 protein to 20 μg/mL with pure water.
2. Prepare substrate mixture: Mix 100 μL of 1 M Tris (pH 8.5), 50 μL of 2 M KCl, 50 μL of 20 mM β-NAD, 50 μL of 40 mM DTT, 50 μL of 200 mM propionaldehyde, and 200 μL of pure water.
3. Add 50 μL of 20 μg/mL Human ALDH1A1 protein to a 96-well plate, then add 50 μL of substrate mixture to initiate the reaction. The blank group contains substrate mixture without the target protein.
4. Zero the microplate reader using the OD value of the blank group, and read the absorbance at 339 nm for 5 min in kinetic mode.
5. Calculate the specific activity:

Human

E. coli

N-6*His

P00352 (M1-S501)

216  [NCBI]

Full Length

MSSSGTPDLPVLLTDLKIQYTKIFINNEWHDSVSGKKFPVFNPATEEELCQVEEGDKEDVDKAVKAARQAFQIGSPWRTMDASERGRLLYKLADLIERDRLLLATMESMNGGKLYSNAYLNDLAGCIKTLRYCAGWADKIQGRTIPIDGNFFTYTRHEPIGVCGQIIPWNFPLVMLIWKIGPALSCGNTVVVKPAEQTPLTALHVASLIKEAGFPPGVVNIVPGYGPTAGAAISSHMDIDKVAFTGSTEVGKLIKEAAGKSNLKRVTLELGGKSPCIVLADADLDNAVEFAHHGVFYHQGQCCIAASRIFVEESIYDEFVRRSVERAKKYILGNPLTPGVTQGPQIDKEQYDKILDLIESGKKEGAKLECGGGPWGNKGYFVQPTVFSNVTDEMRIAKEEIFGPVQQIMKFKSLDDVIKRANNTFYGLSAGVFTKDIDKAITISSALQAGTVWVNCYGVVSAQCPFGGFKMSGNGRELGEYGFHEYTEVKTVTVKISQKNS

Approximately 57 kDa

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

溶液.

Supplied as a 0.22 μm filter solution of 20 mM Tris-HCl, 150 mM NaCl, 20% Glycerol, 1 mM DTT, pH 8.0 or 50 mM Tris-HCl, 300 mM NaCl, pH 7.4, 20% Glycerol, 1 mM Dithiothreitol.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.

• [1]. Calleja LF, et al. Omeprazole as a potent activator of human cytosolic aldehyde dehydrogenase ALDH1A1. Biochim Biophys Acta Gen Subj. 2020 Jan;1864(1):129451.  [Content Brief]