| Cat. No. : |
CYP1A2-121H |
| Description : |
This gene encodes a member of the cytochrome P450 superfamily of enzymes. The cytochrome P450 proteins are monooxygenases which catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids and other lipids. The protein encoded by this gene localizes to the endoplasmic reticulum and its expression is induced by some polycyclic aromatic hydrocarbons (PAHs), some of which are found in cigarette smoke. The enzyme"s endogenous substrate is unknown; however, it is able to metabolize some PAHs to carcinogenic intermediates. Other xenobiotic substrates for this enzyme include caffeine, aflatoxin B1, and acetaminophen. The transcript from this gene contains four Alu sequences flanked by direct repeats in the 3" untranslated region. |
| Source : |
Microsomes prepared from insect cells that were infected with baculovirus containing the cDNA for human CYP1A2 and rabbit cytochrome P450 reductase |
| Amount : |
0.5 nmol, based on spectral P450 content |
| Spectral P450Content : |
1.1 nmol/mL |
| Total Protein Concentration : |
8.66 mg/mL as determined with the BCA assay using BSA as the standard |
| Storagebuffer : |
100 mM potassium phosphate (pH 7.4), 20% glycerol, 1 mM DTT and 0.1 mM EDTA |
| Cytochrome P450 ReductaseActivity : |
1.82 µmol of cytochrome c reduced per minute per milligram of protein |
| PhenacetinO-Deethylase Activity : |
5800 pmol/min/mg of protein. Phenacetin O-deethylase reactions were performed for 10 minutes at 37°C. Reactions contained 0.1 M potassium phosphate (pH 7.4), 10 pmol CYP1A2, 1.3 mM NADP+, 3.33 mM glucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, 3.33 mM magnesium chloride and 0.2 mM phenacetin. The reactions were stopped by the addition of 0.2 volumes of acetonitrile and were centrifuged (10,000 × g) for ten minutes. 40 µL of supernatant was injected onto a 4.5 mm × 70 mm 3 µm C18 HPLC column. |
| HPLC conditions : |
Solvent A: 10% methanol, Solvent B: 25% methanol. Initial conditions: 100% solvent A, after one minute start gradient to 100% solvent B over 6 minutes. The flow rate was 1.0 mL/min. The product was detected by its absorbance at 244 nm. The amount of acetamidophenol produced was calculated by comparison to a standard curve of authentic acetamidophenol |
| Storage : |
Store at –80°C in aliquots. AVOID FREEZE-THAW CYCLES. |
| Safety Precautions : |
Normal precautions exercised in handling laboratory reagents should be followed. |
| Note : |
The information on this certificate of analysis is lot-specific. |
| OfficialSymbol : |
CYP1A2 |
