Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF

R&D Systems | Catalog # 5869-DH

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1 (5869-DH)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P00352

Structure / Form

Noncovalently-linked homotetramer

Applications

Enzyme Activity
View all Aldehyde Dehydrogenase 1-A1/ALDH1A1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Aldehyde Dehydrogenase 1-A1/ALDH1A1 protein
Ser2-Ser501, with an N-terminal Met and 6-His tag

Purity

>90%, 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.

N-terminal Sequence Analysis

Ser2

Predicted Molecular Mass

56 kDa

SDS-PAGE

57 kDa, reducing conditions

Activity

Measured by its ability to produce NADH during the oxidation of propionaldehyde.
The specific activity is >150 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5869-DH
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
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.

Background: Aldehyde Dehydrogenase 1-A1/ALDH1A1

Aldehyde dehydrogenases (ALDHs) are NAD(P)+-dependent enzymes that detoxify aldehydes by oxidizing them to carboxylic acids. Nineteen ALDHs are present in humans, expressed in a variety of organelles and having different substrate preferences (1). ALDH1A1 is a cytosolic enzyme that preferentially oxidizes retinaldehyde to retinoic acid (2). ALDH1A1 is expressed in the epithelium of many organs, including brain, liver, testis, eye lens and cornea (3). ALDH1A1 is highly expressed in brain dopaminergic neurons, where it produces the retinoic acid required for their differentiation and development (4). The retinoic acid produced by ALDH1A1 is also important for the differentiation of hematopoietic stem cells (5). ALDH1A1 is a major enzyme in the oxidation of acetaldehyde, a toxic metabolite of ethanol (6).

References

  1. Marchitti, S.A. et al. (2008) Expert Opin. Drug Metab. Toxicol. 4:697.
  2. Zhao, D. et al. (1996) Eur. J. Biochem. 240:15.
  3. King, G. and Holmes, R. (1997) Adv. Exp. Med. Biol. 414:19.
  4. Jacobs, F.M. et al. (2007) Development 134:2673.
  5. Chute, J.P. et al. (2006) Proc. Natl. Acad. Sci. USA. 103:11707.
  6. Ueshima, Y. et al. (1993) Alcohol Alcohol. 1B:15.

Alternate Names

ALDC, Aldehyde Dehydrogenase 1A1, ALDH-E1, ALDH1A1, PUMB1, RALDH1

Entrez Gene IDs

216 (Human)

Gene Symbol

ALDH1A1

UniProt

P00352

Additional Aldehyde Dehydrogenase 1-A1/ALDH1A1 Products

Product Documents for Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF

For research use only

Citations for Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF

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Protocols

View specific protocols for Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF (5869-DH):

Materials
  • Assay Diluent: deionized water
  • Recombinant Human Aldehyde Dehydrogenase 1‑A1/ALDH1A1 (rhALDH1-A1) (Catalog # 5869-DH)
  • 1 M Tris, pH 8.5
  • 2 M KCl in deionized water
  • DTT, 1M stock in deionized water
  • Propionaldehyde (Sigma, Catalog # 538124), 200 mM stock in deionized water
  • beta -Nicotinamide adenine dinucleotide ( beta -NAD) (Sigma, Catalog # N6522), 100 mM stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Prepare the Substrate Mixture:
    1. 100 µL 1 M Tris, pH 8.5
    2. 50 µL 2 M KCl
    3. 50 µL 20 mM beta -NAD
    4. 50 µL 40 mM DTT
    5. 50 µL 200 mM Propionaldehyde
    6. 200 µL deionized water
      Note: this is enough to assay nine wells. If more volume is needed multiply each component’s volume by the same number to the the desired amount.
  2. Dilute rhALDH1-A1 to 20 µg/mL in Assay Diluent.
  3. Load in a plate 50 μL of 20 µg/mL rhALDH1-A1, and start the reaction by adding 50 μL of Substrate Mixture.
    4. Include a Substrate Blank containing 50 μL of Assay Diluent and 50 μL of Substrate Mixture.
  4. Read at 339 nm in kinetic mode for 5 minutes.

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (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 6220 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhALDH1-A1: 1.0 µg
  • beta -NAD: 1 mM
  • Propionaldehyde: 10 mM

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