Human Cytochromes P450

Human P450s -- Lymphoblast Expressed

CYP1A | CYP1B | CYP2A | CYP2B | CYP2C | CYP2D | CYP2E | CYP2J |CYP3A |CYP4A | CYP4F | CYP19  

NADPH Regenerating System
Cat. No. 451200 and 451220 (formerly B200 and B220 respectively)
NADPH is a necessary cofactor in many xenobiotic metabolism reactions. NADPH is required for the measurement of oxidase activity catalyzed by P450s, FMOs, NADPH-P450 reductase, and many other oxidase enzymes. A common source of NADPH in an oxidase enzyme assay is an NADPH regenerating system that generates NADPH in situ using an enzymatic reaction. For example, glucose-6-phosphate dehydrogenase (G6PDH) will convert NADP+ to NADPH in the presence of the substrate glucose-6-phosphate (Glc-6-PO4).

The BD Biosciences NADPH regenerating system consists of two reagents, Solution A (NADP+ and Glc-6-PO4) and Solution B (G6PDH). Each reagent is sold separately. Combined, these two reagents form an NADPH regenerating system that can be used for all NADPH requiring oxidase assays (cDNA-expressed enzymes and liver fractions). At least 200-400 enzyme assays can be performed using one vial each of Solution A and B. The total number of assays that can be performed is dependent on a researcher's experimental design.
Solution A (Cat. No. 451220 formerly [B220]) Sample Product Insert
Solution B (Cat. No. 451200 formerly [B200]) Sample Product Insert

Human CYP19 (Aromatase), BD Supersomes™ Enzyme
Cat. No. 456260 (formerly P260)
CYP19 catalyzes the conversion of C19 steroids to estrogens and consequently, is a major therapeutic target. Baculovirus-insect cell expressed.

• With cDNA-expressed human P450 reductase
• Available size: 0.5 nmole in 0.5 ml. Representative total protein concentration: 4.0 mg/ml
• Catalytic assay: Aromatase activity
• Representative activity: 1200 pmol product/(min x mg protein), 5 pmol product/pmol P450/min
  

Sample Product Insert
Posters
CYP19 (Aromatase): Characterization of the Recombinant Enzyme and Its Role in the Biotransformation of Xenobiotics
A High Throughput Screen to Identify Inhibitors of Aromatase (CYP19)

Human CYP1A1, BD Supersomes™ Enzyme
Cat. No. 456211 (formerly P211)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml. Representative total protein concentration: 5 mg/ml.
• Representative P450 content: 120 pmole/mg. Catalytic assay: 7-ethoxyresorufin O-deethylase,
• Representative catalytic activity: 4,000 pmole/(mg x min)

Sample Product Insert

Human CYP1A2, BD Supersomes™ Enzyme
Cat. No. 456203 (formerly P203)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 5 mg/ml. Representative P450 content: 200 pmole/mg
• Catalytic assay: 7-ethoxyresorufin O-deethylase
• Representative catalytic activity: 1,200 pmole/(mg x min)
  

Sample Product Insert

Human CYP1B1, BD Supersomes™ Enzyme
Cat. No. 456220 (formerly P220)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 4.5 mg/ml
• Representative P450 content: 220 pmole/mg
• Catalytic assay: 7-ethoxyresorufin-O-deethylase
• Representative catalytic activity: 670 pmole/(mg x min)
  

Sample Product Insert

Human CYP2A6, BD Supersomes™ Enzyme
Cat. No. 456204 (formerly P204)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1.0 nmole P450 in 0.5 ml
• Representative total protein concentration: 5.0 mg/ml
• Representative P450 content: 400 pmole/mg
• Catalytic assay: Coumarin 7-hydroxylase
• Representative catalytic activity: 2,150 pmole/(mg x min)
  

Sample Product Insert

Human CYP2A6+OR+b5, BD Supersomes™ Enzyme
Cat. No. 456254 (formerly P254)
This product is baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and human cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 5.3 mg/ml
• Representative P450 concentration: 190 pmole/mg
• Representative cytochrome b5 concentration: 470 pmole/mg
• Representative catalytic assay: coumarin 7-hydroxylase activity, 4,200 pmol/(mg x min), 22 pmol/pmolP450/min
  

Sample Product Insert

Human CYP2B6, BD Supersomes™ Enzyme
Cat. No. 456210 (formerly P210)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 6 mg/ml
• Representative P450 content: 150 pmole/mg
• Catalytic assay: 7-ethoxytrifluoromethylcoumarin O-deethylase
• Representative catalytic activity: 340 pmole/(mg x min)
  

Sample Product Insert

Human CYP2B6+OR+b5, BD Supersomes™ Enzyme
Cat. No. 456255 (formerly P255)
This product is baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and human cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 7.8 mg/ml
• Representative P450 concentration: 130 pmole/mg
• Representative cytochrome b5 concentration: 280 pmole/mg
• Representative catalytic assay: 7-ethoxy-4-trifluoromethylcoumarin deethylase activity, 1,400 pmol/(mg x min), 11 pmol/pmolP450/min
  

Sample Product Insert

Human CYP2C8, BD Supersomes™ Enzyme
Cat. No. 456212 (formerly P212)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1 nmole P450 in 0.5 ml
• Representative total protein concentration: 5 mg/ml
• Representative P450 content: 400 pmole/mg
• Catalytic assay: 6a-paclitaxel hydroxylase
• Representative catalytic activity: 1,400 pmole/(mg x min)
  

Sample Product Insert

Human CYP2C8 + b5, BD Supersomes™ Enzyme
Cat. No. 456252 (formerly P252)
Baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and cytochrome b5. The incorporation of cytochrome b5 in these microsomes increases the cytochrome P450 activity.

• Available size: 0.5 nmole P450 in 0.5 ml.
• Representative total protein concentration: 3.0 mg/ml.
• Representative P450 content: 370 pmole/mg.
• Representative b5 content: 590 pmole/mg.
• Catalytic assay: paclitaxel-6(alpha)-hydroxylase activity
• Representative catalytic activity: 2,200 pmole/(mg x min)
  

Sample Product Insert

Human CYP2C9*1, BD Supersomes™ Enzyme
Cat. No. 456218 (formerly P218)
This is the wild type CYP2C9. Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1 nmole P450 in 0.5 ml
• Representative total protein concentration: 4 mg/ml. Representative P450 content: 500 pmole/mg
• Catalytic assay: diclofenac 4'-hydroxylase
• Representative catalytic activity: 8,000 pmole/(mg x min)
  

Sample Product Insert

Human CYP2C9*1 + b5, BD Supersomes™ Enzyme
Cat. No. 456258 (formerly P258)
This is the wild type CYP2C9.Baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and cytochrome b5. The incorporation of cytochrome b5 in these microsomes increases the cytochrome P450 activity.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 2.5 mg/ml
• Representative P450 content: 380 pmole/mg
• Representative b5 content: 700 pmole/mg
• Catalytic assay: diclofenac-4'-hydroxylase activity
• Representative catalytic activity: 11,000 pmole/(mg x min)

Note: While we have found the presence of b5 to increase diclofenac 4'-hydroxylation, only ~20%, (S)-warfarin and 7-ethoxy-3-cyanocoumarin metabolism were found to increase four-fold.
Sample Product Insert

Human CYP2C9*2, BD Supersomes™ Enzyme
Cat. No. 456209 (formerly P209)
CYP2C9*2 (Cys144) is an allelic varient of CYP2C9. The frequency of this allele is reported to beapproximately 10%. Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1 nmole P450 in 0.5 ml
• Representative total protein concentration: 3 mg/ml
• Representative P450 content: 600 pmole/mg
• Catalytic assay: diclofenac 4'-hydroxylase
• Representative catalytic activity: 12,000 pmole/(mg x min)
  

Sample Product Insert

Human CYP2C9*3, BD Supersomes™ Enzyme
Cat. No. 456242 (formerly P242)
CYP2C9*3 (359-Leu) is an allelic varient of CYP2C9. The frequency of this allele is reported to be 0.06, 0.005, and 0.026 in a Caucasian-American, African-American, and Chinese-Taiwanese population. Amino acid 359 lies within a putative, distal substrate binding domain for the CYP2C isoforms. The intrinsic clearance (Vmax/Km) for CYP2C9*3 (456242 [formerly P242]) (S)-warfarin 7-hydroxylase activity was found to be 5.5 fold lower than that for CYP2C9*1 (456218 [formerly P218]). The intrinsic clearance (Vmax/Km) for CYP2C9*3 (456242 [formerly P242]) diclofenac 4'-hydroxylase activity was found to be 1.8 fold lower than that for CYP2C9*1 (456218 [formerly P218]). Similar differences in intrinsic clearance between these two CYP2C9 alleles have been reported by other laboratories. Baculovirus-insect cell expressed with supplemental cDNA expressed human reductase.

• Available size: 1.0 nmole P450 in 0.5 ml
• Representative total protein concentration: 3.0 mg/ml
• Representative P450 concentration: 650 pmole/mg
• Catalytic assay: diclofenac 4'hydroxylase
• Representative catalytic activity, 11,400 pmol/(mg x min)
  

Sample Product Insert

Human CYP2C18, BD Supersomes™ Enzyme
Cat. No. 456222 (formerly P222)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml.
• Representative total protein concentration: 5.0 mg/ml.
• Representative P450 content: 190 pmole/mg.
• Catalytic assay: Diclofenac 4'-hydroxylase, two diclofenac metabolites are produced by CYP2C18, the 4'-hydroxydiclofenac metabolite is the minor of the two metabolites and is quantitated because of the availability of authentic standard
• Representative catalytic activity: 120 pmole/(mg x min)

Note: We have found this enzyme to also 4'-hydroxylate (S)-warfarin.
Sample Product Insert

Human CYP2C19, BD Supersomes™ Enzyme
Cat. No. 456219 (formerly P219)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 3 mg/ml
• Representative P450 content: 360 pmole/mg
• Catalytic assay: (S)-mephenytoin 4'-hydroxylase
• Representative catalytic activity: 900 pmole/(mg x min)
  

Sample Product Insert

Human CYP2C19 + b5, BD Supersomes™ Enzyme
Cat. No. 456259 (formerly P259)
Baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and cytochrome b5. The incorporation of cytochrome b5 in these microsomes increases the cytochrome P450 activity.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 3.0 mg/ml
• Representative P450 content: 380 pmole/mg
• Representative b5 content: 580 pmole/mg
• Catalytic assay: (S)-mephenytoin-4'-hydroxylase
• Representative catalytic activity: 16,500 pmole/(mg x min)
  

Sample Product Insert

Human CYP2D6*1, BD Supersomes™ Enzyme
Cat. No. 456217 (formerly P217)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 5 mg/ml
• Representative P450 content: 250 pmole/mg
• Catalytic assay: bufuralol 1'-hydroxlase
• Representative catalytic activity: 9,200 pmole/(mg x min)
  

Sample Product Insert

Human CYP2D6*10 Allelic Variant Kit
Cat. No. 456240 (formerly P240)
CYP2D6 is polymorphic (either null alleles or amino acid substitution(s)) in most human populations. The CYP2D6*10 allele contains two amino acid substitutions, P34S and S486T. This allele is quite common in Asians and has been linked to reduced rates of metabolism of known CYP2D6 substrates. In this kit CYP2D6*10 and CYP2D6*1 are each co-expressed with human cytochrome P450 oxidoreductase (OR) in insect cells using a baculovirus expression vector. Both alleles contain similar reductase to P450 ratios. In validation experiments CYP2D6*10 exhibited 25-fold and 12-fold lower in vitro intrinsic clearances, relative to CYP2D6*1, for bufuralol 1'-hydroxylation and debrisoquine 4-hydroxylation respectively.

Kit Contents:

• Human CYP2D6*10 BD Supersomes™ Enzyme
• Human CYP2D6*1 BD Supersomes™ Enzyme
• Insect control microsomes.
  

Sample Product Insert

Human CYP2E1 + b5, BD Supersomes™ Enzyme
Cat. No. 456206 (formerly P206)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase and cytochrome b5.

• Available size: 1.0 nmole P450 in 0.5 ml
• Representative total protein concentration: 4.0 mg/ml
• Representative P450 content: 490 pmole/mg
• Catalytic assay: p-Nirophenol hydroxylase
• Representative catalytic activity: 6,500 pmole/(mg x min)
  

Sample Product Insert

Human CYP2J2 + b5, BD Supersomes™ Enzyme
Cat. No. 456264
CYP2J2 is expressed at high levels in cardiac tissue. It is expressed at lower levels in the intestine, liver, kidney, and lung. CYP2J2 is active for the oxidation of arachidonic acid to epoxyeicosatrienoic acids (EETs), which are important for heart and kidney function. CYP2J2 has also been shown to metabolize xenobiotics such as diclofenac and bufuralol¹. CYP2J2 is the predominant P450 for the hydroxylation of ebastine and astemizole (non-sedating anti-histamines) in human intestinal microsomes². More recent studies have shown that CYP2J2 is a high affinity enzyme for the hydroxylation of terfenadine³. Baculovirus-insect cell expressed with supplemental cDNA expressed reductase and cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 4.0 mg/ml
• Representative P450 content: 210 pmole/mg
• Catalytic assay: Terfenadine Hydroxylase
• Representative catalytic activity: 3,800 pmole/(mg x min)
  

1. Scarborough, P., et.al., Drug Met. Rev., 31:205-234, 1999
2. Hashizume, T. et.al., JPET, 300:298-304, 2002, Matsumoto, S., et.al., DMD, 30:1240-1245, 2002
3. European ISSX, 2003, abstract No. 242

Sample Product Insert

Human CYP3A4, BD Supersomes™ Enzyme
Cat. No. 456207 (formerly P207)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1 nmole P450 in 0.5 ml
• Representative total protein concentration: 5 mg/ml
• Representative P450 content: 400 pmole/mg
• Catalytic assay: testosterone 6b-hydroxylase
• Representative catalytic activity: 2,200 pmole/(mg x min)
  

Sample Product Insert

Human CYP3A4 + b5, BD Supersomes™ Enzyme
Cat. No. 456202 (formerly P202)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase and cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 4 mg/ml
• Representative P450 content: 300 pmole/mg
• Catalytic assay: testosterone 6b-hydroxylase
• Representative catalytic activity: 32,000 pmole/(mg x min)
  

Sample Product Insert

Human CYP3A5, BD Supersomes™ Enzyme
Cat. No. 456235 (formerly P235)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 1 nmole P450 in 0.5 ml
• Representative total protein concentration: 7 mg/ml
• Representative P450 content: 250 pmole/mg
• Catalytic assay: testosterone 6b-hydroxylase
• Representative catalytic activity: 900 pmole/(mg x min)
  

Sample Product Insert

Human CYP3A5 + b5, BD Supersomes™ Enzyme
Cat. No. 456256
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase and cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 4 mg/ml
• Representative P450 content: 200 pmole/mg
• Catalytic assay: testosterone 6b-hydroxylase
• Representative catalytic activity: 25,000 pmole/(mg x min)

Sample Product Insert

Human CYP3A7+OR+b5, BD Supersomes™ Enzyme
Cat. No. 456237 (formerly P237)
CYP3A7 is expressed in fetal liver and adult kidney. This product is baculovirus-insect cell expressed with supplemental cDNA expressed human reductase and human cytochrome b5.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 5 mg/ml
• Representative P450 concentration: 180 pmole/mg
• Representative cytochrome b5 concentration: 1040 pmole/mg
• Representative catalytic assay: testosterone 6(beta)-hydroxylase activity, 210 pmol/(mg x min), 1.1 pmol/pmolP450/min

This product is expressed in Sf9 insect cells and therefore Sf9 insect control microsomes (Cat. No. 456299 [formerly P299]) should be used as a control for native activities.
Sample Product Insert

Human CYP4A11, BD Supersomes™ Enzyme
Cat. No. 456221 (formerly P221)
Baculovirus-insect cell expressed with supplemental cDNA expressed reductase.

• Available size: 0.5 nmole P450 in 0.5 ml
• Representative total protein concentration: 6 mg/ml
• Representative P450 content: 150 pmole/mg
• Catalytic assay: lauric acid omega-hydroxlase
• Representative catalytic activity: 7,000 pmole/(mg x min)
  

Sample Product Insert

Human CYP4F2, BD Supersomes™ Enzyme
Cat. No. 456272 (formerly P272)
Cytochrome P450 4F2 has recently been identified in human liver. CYP4F2 possess a high specificity for LTB4 20-OH hydroxylation. However, the Km values for LTB4 metabolism by hepatic CYP4F enzymes are reported to be several fold higher than the value reported for CYP4F3A. The role of this enzyme in xenobiotic metabolism is currently being defined. This product is baculovirus-insect cell expressed and also contains cDNA-expressed human P450 reductase and human cytochrome b5.

• Available size: 0.5 ml
• Representative total protein concentration: 4 mg/ml. Catalytic assay: 20-hydroxy leukotriene B4 activity
• Representative activity: 500 pmol product/(min x mg protein), 2 pmol product/pmol P450/min
  

Sample Product Insert

Human CYP4F3A, BD Supersomes™ Enzyme
Cat. No. 456273 (formerly P273)
Cytochrome P450 4F3A (CYP4F3A) is a major enzyme involved in the w-oxidation of LTB4 in humans, yielding the biologically less active 20-OH product. CYP4F3A is a non-hepatic P450 enzyme; its expression is localized to the ER of polymorphonuclear leukocytes (PMNL). Baculovirus expressed CYP4F3A, and PMNL, possess a similar low Km (less than 1 uM) for the formation of 20-OH LTB4. The role of this enzyme in xenobiotic metabolism is currently being defined. This product is baculovirus-insect cell expressed and also contains cDNA-expressed human P450 reductase and human cytochrome b5.

• Available size: 0.5 ml
• Representative total protein concentration: 14 mg/ml
• Catalytic assay: 20-hydroxy leukotriene B4 activity
• Representative activity: 2,000 pmol product/(min x mg protein), 56 pmol product/pmol P450/min
  

Sample Product Insert

Human CYP4F3B, BD Supersomes™ Enzyme
Cat. No. 456274 (formerly P274)
Cytochrome P450 4F3B has recently been identified in human liver. CYP4F3B possess a high specificity for LTB4 20-OH hydroxylation. However, the Km values for LTB4 metabolism by hepatic CYP4F enzymes are reported to be several fold higher than the value reported for CYP4F3A. The role of this enzyme in xenobiotic metabolism is currently being defined. This product is baculovirus-insect cell expressed and also contains cDNA-expressed human P450 reductase and human cytochrome b5.

• Available size: 0.5 ml
• Representative total protein concentration: 3 mg/ml
• Catalytic assay: 20-hydroxy leukotriene B4 activity
• Representative activity: 2500 pmol product/(min x mg protein), 7.4 pmol product/pmol P450/min
  

Sample Product Insert

Human CYP4F12, BD Supersomes™ Enzyme
Cat. No. 456275
CYP4F12 is expressed in both the liver and the intestine. CYP4F12 has been shown to be active for the hydroxylation of ebastine¹, and terfenadine². This product is baculovirus-insect cell expressed and also contains cDNA-expressed human P450 reductase and human cytochrome b5.

• Available size: 0.5 ml
• Representative total protein concentration: 4mg/ml
• Catalytic assay: Terfenadine Hydroxylase activity
• Representative activity: 2100 pmol product/(min x mg protein), 9.9 pmol product/pmol P450/min
  

1. Hashizume, T. et.al., JPET, 300:298-304, 2002
2. European ISSX, 2003, abstract No. 242

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