Recombinant Human POR Protein, CF

R&D Systems | Catalog # 6340-PR

R&D Systems
Discontinued Product
6340-PR has been discontinued. An alternative/replacement product is available: 6340-PRB. View all POR/Cytochrome P450 Reductase products.

Key Product Details

Source

Sf 21 (baculovirus)

Accession Number

P16435.2

Applications

Enzyme Activity
View all POR/Cytochrome P450 Reductase Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human POR/Cytochrome P450 Reductase protein
Arg45-Ser677, with N-terminal amino acids WAGALA and 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

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

N-terminal Sequence Analysis

N-terminal Trp & P16435.2 Ser62

Predicted Molecular Mass

73 kDa & 70 kDa

SDS-PAGE

65-80 kDa, reducing conditions

Activity

Measured by the reduction of cytochrome c using NADPH as the cofactor.
The specific activity is >6,500 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 6340-PR in the following applications:

Formulation, Preparation, and Storage

6340-PR
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.

Background: POR/Cytochrome P450 Reductase

NADPH-Cytochrome P450 Reductase (P450R) is an essential component of the cytochrome P450 monooxygenase system of eukaryotic cells (1). P450R is anchored in the endoplasmic reticulum membrane with its catalytic domain residing in the cytosol. P450R is a flavoprotein, containing one molecule each of FMN and FAD, which are essential for the transfer of electrons from NADPH to the cytochromes P450 (2). This reduction is necessary for cytochromes P450 to perform each cycle of oxidation. P450R is also capable of transferring electrons to cytochrome b5, heme oxygenase, the fatty acid elongation system, and other proteins. Mutations of P450R can result in disordered steroidogenesis and Antley-Bixler syndrome.

References

  1. Philips, A.H. and R.G. Langdon (1962) J. Biol. Chem. 237:2652.
  2. Iyanagi, T. and H.S. Mason (1973) Biochemistry 12:2291.
  3. Flueck C.E. et al. (2004) Nat. Genet. 36:228.

Long Name

NADPH Cytochrome P450 Reductase

Alternate Names

CPR, CYPOR, Cytochrome P450 Reductase, P450R

Entrez Gene IDs

5447 (Human)

Gene Symbol

POR

UniProt

P16435.2

Additional POR/Cytochrome P450 Reductase Products

Product Documents for Recombinant Human POR Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human POR Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Redox Enzymes

Customer Reviews for Recombinant Human POR Protein, CF (1)

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Protocols

View specific protocols for Recombinant Human POR Protein, CF (6340-PR):

Materials
  • Assay Buffer: 0.3 M KH2PO4, pH 8.0
  • Recombinant Human POR/Cytochrome P450 Reductase (rhPOR) (Catalog # 6340-PR)
  • Substrate: beta -NADPH (Sigma, Catalog # N7505), 10 mM stock in deionized water (8.33 mg/mL)
  • Cytochrome C, Bovine heart (Sigma, Catalog # C3131), 2 mg/mL stock in deionized water
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhPOR to 0.2 ng/μL in Assay Buffer.
  2. Combine 480 μL of Assay Buffer, 500 μL of 2 mg/mL Cytochrome C, and 20 μL of 10 mM beta -NADPH.
  3. Load 50 μL of 0.2 ng/μL rhPOR and start the reaction by adding 50 μL of Substrate Mixture. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL Substrate Mixture.
  4. Read in kinetic mode for 6 minutes with 1 minute lag time at an absorbance of 550 nm.
  5. Calculate specific activity:

     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 21100 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhPOR: 0.010 μg
  • Cytochrome C: 0.5 mg/ml
  • beta -NADPH: 0.1 mM

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