rev: 5/16/2008

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CYTOCHROME P450

NATIVE ENZYMES 

CYP450 2A6

NATIVE  HUMAN LIVER CYP2A6

PRODUCT NUMBER :    RDI-CYP2A6         $855.75/nmol

P450 Enzyme Purified from Human Liver Microsomes


LOT #3

P450 CONTENT = 3.9 nmol/ml

PROTEIN CONTENT = 0.64 mg/ml

SPECIFIC CONTENT = 6.1 nmol P450/mg protein


CYP2A6 was purified from liver microsomes from a single human subject using conventional techniques, including hydrophobic, anion-exchange, and hydroxylapatite adsorption chromatographies. Human CYP2A6 is provided in a solution containing 100 mM potassium phosphate buffer (pH 7.4), 0.1 mM EDTA, 0.1 mM DTT, and 20% glycerol.

  • Purity

Purity has been determined by electrophoresis on 7.5% acrylamide gels run with the discontinuous buffer system. CYP2A6 migrates as a single band with a molecular weight of 49.5 kDa (see Fig. 1, lane C). CYP2A6 is a low-spin hemeprotein when oxidized with a ferrous carbonyl Soret maximum at 451 nm.

 


Picture of Gel

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

Lanes A & J, liver microsomes (10 µg);

Lane B, CYP2D6 (0.5 µg);

lane C, CYP2A6 (0.5 µg);

Lane D, CYP2C19 (0.5 µg);

Lane E, CYP2C8 (0.5 µg);

Lane F, Molecular Weight Standards (0.5 µg each);

Lane G, CYP2E1 (0.5 µg);

Lane H, CYP2C9 (0.5 µg);

Lane I, CYP4F4 (0.5 µg).

 

 

¨Reconstitution

CYP2A6 catalytic activity is assessed upon reconstitution of the enzyme with NADPH:P450 reductase, synthetic dilauroylphosphatidlycholine and, importantly, cytochrome b5. A reconstituted system containing 50 pmol CYP2A6, 150 pmol human liver P450 reductase, and 15 µg phospholipid exhibits a turnover number of 3.1 min-1 with coumarin as substrate; in the presence of 200 pmol human liver b5, coumarin 7-hydroxylase activity increases to 11.1 min-1. Full details for reconstitution are given in an accompanying instruction sheet.

  • Storage

CYP2A6 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.

 

See also antibodies against: CYP2A6, CYP2C9, CYP2E1,CYP3A4 (in production), CYP2C19 and CYP450 reductase.

 

 

RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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