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Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF

Catalog #: 5869-DH Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
5869-DH-020
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Recombinant Human Aldehyde Dehydrogenase 1-A1/ALDH1A1, CF Summary

Product Specifications

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.
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.
Source
E. coli-derived human Aldehyde Dehydrogenase 1-A1/ALDH1A1 protein
Ser2-Ser501, with an N-terminal Met and 6-His tag
Accession #
P00352
N-terminal Sequence
Analysis
Ser2
Structure / Form
Noncovalently-linked homotetramer
Predicted Molecular Mass
56 kDa
SDS-PAGE
57 kDa, reducing conditions

Product Datasheets

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5869-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.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
216 (Human)
Alternate Names
acetaldehyde dehydrogenase 1; ALDC; aldehyde dehydrogenase 1 family, member A1; aldehyde dehydrogenase 1, soluble; Aldehyde Dehydrogenase 1A1; Aldehyde Dehydrogenase 1-A1; Aldehyde dehydrogenase family 1 member A1; Aldehyde dehydrogenase, cytosolic; aldehyde dehydrogenase, liver cytosolic; ALDH class 1; ALDH1; ALDH11; ALDH1A1; ALDH-E1; ALHDII; EC 1.2.1; EC 1.2.1.36; MGC2318; PUMB1; RALDH 1; RALDH1; retinal dehydrogenase 1; retinaldehyde dehydrogenase 1

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