ALDH1A1 Protein, 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.).

Aldehyde Dehydrogenase 1-A1 (ALDH1A1) is part of the aldehyde dehydrogenases family. Human ALDH1A1 is a cytosolic homotetramer expressed in several tissues (skin, eyes, liver, kidney, testis and brain). ALDH1A1oxidizes a broad range of aliphatic and aromatic aldehydes including those formed endogenously such as retinalsand 3,4-dihydroxyphenylacetaldehyde (DOPAL), xenobiotics such as benzaldehyde or those produced during lipid peroxidation^[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.

Solution.

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]