Recombinant Human Aldehyde Dehydrogenase 3-A1/ALDH3A1, CF

Catalog #: 6705-DH Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
6705-DH-020

R&D Systems Recombinant Proteins and Enzymes

1 Image

All Aldehyde Dehydrogenase 3-A1/ALDH3A1 Proteins and Enzymes

Product Details

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Aldehyde Dehydrogenase 3-A1/ALDH3A1, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

Measured by the ability to catalyze the oxidation of 4-nitrobenzaldehyde. The specific activity is >6000 pmol/min/μg, as measured under the described conditions.

Source

E. coli-derived human Aldehyde Dehydrogenase 3-A1/ALDH3A1 protein
Ser2-His453, with an N-terminal Met and 6-His tag

Accession #

P30838

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

51 kDa

SDS-PAGE

51-56 kDa, reducing conditions

Product Datasheets

6705-DH

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

6705-DH

Formulation	Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -70 °C as supplied. 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials

Assay Buffer: 50 mM Tris, 5 mM DTT, pH 9.0
Recombinant Human Aldehyde Dehydrogenase 3-A1/ALDH3A1 (rhALDH3A1) (Catalog # 6705-DH)
Nicotinamide adenine dinucleotide phosphate (NADP⁺) (Sigma, Catalog # N5755), 50 mM stock in deionized water
4-Nitrobenzaldehyde (4-NBA) (Sigma, Catalog # 72800), 200 mM stock in DMSO
96-well Clear Plate (Costar, Catalog # 92592)
Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent

Dilute rhALDH3A1 to 4 ng/µL in Assay Buffer.
Dilute NADP⁺ to 2 mM in Assay Buffer.
Dilute 4-NBA to 4 mM in Assay Buffer.
Form Substrate Mixture by combining equal volumes of 2 mM NADP⁺ and 4 mM 4-NBA.
Load 50 µL of the 4 ng/µL rhALDH3A1 into the plate. Include a Substrate Blank containing 50 µL of Assay Buffer.
Start the reaction by adding 50 µL of Substrate Mixture to the wells.
Read plate at 340 nm (absorbance) in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (OD/min) x well volume (L) x 10¹² pmol/mol
	ext. coeff (M^-1cm^-1) x path corr.* (cm) x amount of enzyme (µg)

      *Adjusted for Substrate Blank
     **Using the extinction coefficient 6270 M^-1cm^-1     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:

rhALDH3A1: 0.2 µg
NADP⁺: 0.5 mM
4-NBA: 1 mM

Reconstitution Calculator

Background: Aldehyde Dehydrogenase 3-A1/ALDH3A1

Aldehyde Dehydrogenases (ALDHs) are NAD(P)⁺-dependent enzymes that catalyze the oxidation of endogeneously produced and exogeneous aldehydes to their corresponding acids (1). They are involved in the detoxification of alcohol-derived acetaldehyde and in the metabolism of corticosteroids, biogenic amines, neurotransmitters, and in lipid peroxidation. ALDH3A1 is also known as stomach aldehyde dehydrogenase. It exists as a homodimer, and prefers NADP⁺ over NAD⁺ as its co‑factor (2). It preferentially oxidizes aromatic and medium-chain (6 carbons or more) saturated and unsaturated aldehyde substrates (3). The enzyme is highly expressed in stomach and cornea (2, 4). In the cornea, its proposed roles have been to absorb UV‑light, reduce oxidative damage, maintain corneal refractive and transparence properties, and display chaperone-like activity (4, 5). It also has been identified as a lung cancer biomarker (6).

References

Marchitti, S. A. et al. (2007) Pharmacol. Rev. 59:125.
Yin, S. J. et al. (1991) FEBS Lett. 283:85.
Hsu, L. C. et al. (1992) J. Biol. Chem. 267:3030.
Estey, T. et al. (2007) Exp. Eye Res. 84:3.
Estey, T. et al. (2010) PLOS One. 5:e15218.
Kim, B. et al. (2007) Cancer Res. 67:7431.

Entrez Gene IDs

218 (Human); 11670 (Mouse); 25375 (Rat)

Alternate Names

aldehyde dehydrogenase 3 family, member A1; Aldehyde dehydrogenase 3; Aldehyde Dehydrogenase 3A1; Aldehyde Dehydrogenase 3-A1; Aldehyde dehydrogenase family 3 member A1; aldehyde dehydrogenase isozyme 3; aldehyde dehydrogenase type III; ALDH3A1; ALDH3aldehyde dehydrogenase, dimeric NADP-preferring; ALDHIII; EC 1.2.1; EC 1.2.1.5; MGC10406; stomach aldehyde dehydrogenase