rev:April 9, 1997

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CYTOCHROME P450

NATIVE ENZYMES 

PRODUCT NUMBER : RDI-P450OXRD  $500.00/nmol

HUMAN LIVER NADPH:P450 OXIDOREDUCTASE
Enzyme Purified from Human Liver Microsomes


LOT # (sample data-see sheet with shipment)

ENZYME ACTIVITY = 57.9   nmol (188,000 units) /ml

PROTEIN CONTENT =  6.88 mg/ml

SPECIFIC ACTIVITY =  8.41 nmol (27,000 units/mg protein


NADPH:P450 oxidoreductase (P450 reductase) was purified from liver microsomes from a single human subject using conventional techniques, including hydrophobic, anion-exchange, affinity, and hydroxylapatite adsorption chromatographies. Human P450 reductase is provided in 100 mM potassium phosphate buffer (pH 7.4), 0.1 mM EDTA, 0.1 mM DTT, and 20% glycerol.

Purity has been determined by electrophoresis on 10% acrylamide gels run with the discontinuous buffer system. P450 reductase migrates as a single band with a molecular weight of 75 kDa (see Fig. 1, lane C). One unit P450 reductase activity is defined as that amount catalyzing reduction of 1 nmol ferricytochrome c/min at 30°C in 300 mM potassium phosphate buffer (pH 7.7). One nmol P450 reductase contains 3,370 units of enzyme activity.




Fig. 1. SDS-PAGE analysis of purified human liver P450 reductase.

Lane A, cytochrome b5 (0.5 µg);

Lane B, liver microsomes (10 µg);

Lane C, P450 Reductase (0.5 µg);










Human P450 reductase serves, in combination with phospholipid, to reconstitute the activity of purified P450 enzymes. The molar ratio of P450 reductase to P450 enzyme used in the reconstituted system should be at least 3:1 [e.g., 150 pmol (ca 500 units) P450 reductase:50 pmol P450]. P450 reductase should be allowed to reconstitute with a given P450 enzyme for 3 min at 37°C in the presence of phospholipid. Full details for reconstitution are given in an accompanying instruction sheet.

P450 reductase should be stored @ -80°C. Avoid repeated freeze-thawing cycles.

Precaution: For in vitro research use only-not for use in or on humans or animals-not for use in diagnostics. Not responsible for any patent infringements with the use or derivation of these products.

RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (800) 370-2222

      or (201) 584-7093  after June 1, 1997:  (973) 584-7093

fax     (978) 371-2266   after June 1, 1997:  (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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