Recombinant Human POR/Cytochrome P450 Reductase Protein, CF

R&D Systems | Catalog # 6340-PRB

His-tag
R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human POR/Cytochrome P450 Reductase Protein (6340-PRB)
  • 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

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
Ser62-Ser677 with an N-terminal Met and 6-His tag

Purity

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

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

71 kDa

SDS-PAGE

63-69 kDa, under reducing conditions.

Activity

Measured by the reduction of cytochrome c using NADPH as the cofactor.
The specific activity is >1800 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human POR/Cytochrome P450 Reductase Protein, CF

Recombinant Human POR/Cytochrome P450 Reductase His-tag Protein Enzyme Activity.

Recombinant Human POR His-tag (Catalog # 6340-PRB) is measured by the reduction of cytochrome c using NADPH as the cofactor.

Recombinant Human POR/Cytochrome P450 Reductase His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human POR/Cytochrome P450 Reductase His-tag Protein (Catalog # 6340-PRB) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 63-69 kDa, under reducing conditions.

Formulation, Preparation, and Storage

6340-PRB
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: POR/Cytochrome P450 Reductase

NADPH-Cytochrome P450 Reductase (P450R), or POR, 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. The protein is composed of four domains: an N-terminal FMN‑binding domain, a connecting domain, and C-terminal FAD- and NADPH-binding domains (2). The flavoprotein P450R contains one molecule each of FMN and FAD, which are essential for the transfer of electrons from NADPH to the P450 cytochromes (3). This reduction is necessary for cytochromes P450 to perform each cycle of oxidation. P450R plays a vital role in metabolism and detoxification of xenobiotic and endobiotic compounds (4). P450R is also capable of transferring electrons to cytochrome b5, heme oxygenase, the fatty acid elongation system, and other proteins (4). Mutations of P450R can result in disordered steroidogenesis and bone defects in Antley-Bixler syndrome (5, 6) and expression of P450R has been correlated with patient outcome in breast cancer (7).

References

  1. Philips, A.H. and R.G. Langdon (1962) J. Biol. Chem. 237:2652.
  2. Wang, M. et al. (1997) Proc. Natl. Acad. Sci. 94:8411.
  3. Iyanagi, T. and H.S. Mason (1973) Biochemistry 12:2291.
  4. Iyanagi, T. (2007) Int. Rev. Cytol. 260:35.
  5. Flueck C.E. et al. (2004) Nat. Genet. 36:228.
  6. Polusani, S.R. et al. (2011) Biochem. Biophys. Res. Commun. 411:490.
  7. Pederson, M.H. et al. (2019) Int. J. Cancer 144:631.

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/Cytochrome P450 Reductase 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.

Product Specific Notices for Recombinant Human POR/Cytochrome P450 Reductase Protein, CF

For research use only

Related Research Areas

Redox Enzymes

Customer Reviews for Recombinant Human POR/Cytochrome P450 Reductase Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human POR/Cytochrome P450 Reductase Protein, CF and earn rewards!

Have you used Recombinant Human POR/Cytochrome P450 Reductase Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human POR/Cytochrome P450 Reductase Protein, CF (6340-PRB):

Materials
  • Assay Buffer: 0.3 M Potassium Phosphate, pH 8.0
  • Recombinant Human POR/Cytochrome P450 Reductase His-tag (rhPOR) (Catalog # 6340-PRB)
  • Substrate: beta -NADPH, 10 mM stock in deionized water
  • Cytochrome C, Bovine heart, 2 mg/mL stock in deionized water
  • Clear 96 well Plate (Catalog # DY990)
  • Plate Reader with Absorbance Read Capability
  1. Dilute rhPOR to 0.2 μg/mL in Assay Buffer.
  2. Prepare a Substrate Mixture containing 200 µM beta -NADPH and 1 mg/mL Cytochrome C in Assay Buffer.
  3. Load 50 μL of 0.2 μg/mL 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: 100 μM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes