CYP3A4 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Cytochrome P450 3A4 (CYP3A4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Cytochrome P450 3A4 (CYP3A4) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

1,4-cineole 2-exo-monooxygenase;1,8-cineole 2-exo-monooxygenase; Albendazole monooxygenase; albendazole monooxygenase (sulfoxide-forming) ; albendazole sulfoxidase; cholesterol 25-hydroxylase; CP33; CP34; CYP3A; CYP3A3; CYPIIIA3; CYPIIIA4; cytochrome P450 3A3; cytochrome P450 3A4; cytochrome P450 HLp; cytochrome P450 NF-25; cytochrome P450, family 3, subfamily A, polypeptide 4; cytochrome P450, subfamily IIIA (niphedipine oxidase), polypeptide 3; cytochrome P450, subfamily IIIA (niphedipine oxidase), polypeptide 4; cytochrome P450-PCN1; glucocorticoid-inducible P450; HLP; NF-25; nifedipine oxidase; P450C3; P450-III, steroid inducible; P450PCN1; quinine 3-monooxygenase; taurochenodeoxycholate 6-alpha-hydroxylase; VDDR3.

Cytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It performs a variety of oxidation reactions (e.g. caffeine 8-oxidation, omeprazole sulphoxidation, midazolam 1'-hydroxylation and midazolam 4-hydroxylation) of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. Acts as a 1,8-cineole 2-exo-monooxygenase. The enzyme also hydroxylates etoposide (PubMed:11159812) . Catalyzes 4-beta-hydroxylation of cholesterol. May catalyze 25-hydroxylation of cholesterol in vitro (PubMed:21576599) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Cytochrome P450 3A4 (CYP3A4) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cytochrome P450 3A4 (CYP3A4) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Cytochrome P450 3A4 (CYP3A4), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Cytochrome P450 3A4 (CYP3A4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Component Amount CYP3A4 Microplate 96 Wells (12 x 8 Well strips) CYP3A4 Lyophilized Standard 2 100X CYP3A4 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Component Amount Component Amount CYP3A4 Microplate 96 Wells (12 x 8 Well strips) CYP3A4 Microplate 96 Wells (12 x 8 Well strips) CYP3A4 Lyophilized Standard 2 CYP3A4 Lyophilized Standard 2 100X CYP3A4 HRP-Detector Anitbody 1 x 120 uL 100X CYP3A4 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Stop Solution 1 x 20 mL