APPLICATION
Application Note
Cytochrome P450 2D6 (CYP2D6), a member of the cytochrome P450 mixed-function oxidase system, is one of the most important enzymes involved in the metabolism of xenobiotics in the body. In particular, CYP2D6 is responsible for the metabolism and elimination of approximately 25% of clinically used drugs, via the addition or removal of certain functional groups-specifically, hydroxylation, demethylation, and dealkylation. CYP2D6 is primarily expressed in the liver. It is also highly expressed in areas of the central nervous system, including the substantia nigra. This enzyme also metabolizes several endogenous substances, such as hydroxytryptamines, neurosteroids, and both m-tyramine and p-tyramine which CYP2D6 metabolizes into dopamine in the brain and liver. Besides, Cytochrome P450 Reductase (CPR) has been identified as an interactor of CYP2D6, thus a binding ELISA assay was conducted to detect the interaction of recombinant human VDR and recombinant human CPR. Briefly, CYP2D6 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to CPR-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-CYP2D6 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of CYP2D6 and CPR was in a dose dependent manner.
Observed MW
27 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Leu236~Gln472 (NP_000097.3)
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
cytochrome P450 family 2 subfamily D member 6 , CPD6 , CYP2D , CYP2D7AP , CYP2D7BP , CYP2D7P2 , CYP2D8P2 , CYP2DL1 , CYPIID6 , P450-DB1 , P450C2D , P450DB1
Background
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. This protein localizes to the endoplasmic reticulum and is known to metabolize as many as 25% of commonly prescribed drugs. Its substrates include antidepressants, antipsychotics, analgesics and antitussives, beta adrenergic blocking agents, antiarrythmics and antiemetics. The gene is highly polymorphic in the human population; certain alleles result in the poor metabolizer phenotype, characterized by a decreased ability to metabolize the enzyme's substrates. Some individuals with the poor metabolizer phenotype have no functional protein since they carry 2 null alleles whereas in other individuals the gene is absent. This gene can vary in copy number and individuals with the ultrarapid metabolizer phenotype can have 3 or more active copies of the gene. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2014]
Database
Research Area
DATA IMAGES
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GTX00199-pro Functional Assay Image
Functional ELISA analysis of GTX00199-pro Human Cytochrome P450 2D6 protein which can bind immobilized CPR protein.
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GTX00199-pro Image
WB analysis of GTX00199-pro Human Cytochrome P450 2D6 protein.
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GTX00199-pro Image
SDS-PAGE analysis of GTX00199-pro Human Cytochrome P450 2D6 protein.
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REFERENCE
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REVIEW
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