CYP7A1 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Cytochrome P450 7A1 (CYP7A1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Cytochrome P450 7A1 (CYP7A1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

24-hydroxycholesterol 7-alpha-hydroxylase; cholesterol 7-alpha-hydroxylase; cholesterol 7-alpha-monooxygenase; CYPVII; CYPVIIc; cytochrome P450 7A1; cytochrome P450, 7a1.

Catalyzes a rate-limiting step in cholesterol catabolism and bile acid biosynthesis by introducing a hydrophilic moiety at position 7 of cholesterol. Important for cholesterol homeostasis.

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 7A1 (CYP7A1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cytochrome P450 7A1 (CYP7A1) . 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 7A1 (CYP7A1), 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 7A1 (CYP7A1) 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

Matrix Recovery range (%) Average (%) Serum (n=5) 85-104 99 EDTA Plasma (n=5) 80-101 90 Heparin Plasma (n=5) 86-99 91 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 85-104 99 Serum (n=5) 85-104 99 EDTA Plasma (n=5) 80-101 90 EDTA Plasma (n=5) 80-101 90 Heparin Plasma (n=5) 86-99 91 Heparin Plasma (n=5) 86-99 91

Component Amount Anti-CYP7A1 Microplate 96 Wells (12 x 8 Well strips) CYP7A1 Lyophilized Standard 2 100X Biotinylated CYP7A1 Detector Antibody 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-CYP7A1 Microplate 96 Wells (12 x 8 Well strips) Anti-CYP7A1 Microplate 96 Wells (12 x 8 Well strips) CYP7A1 Lyophilized Standard 2 CYP7A1 Lyophilized Standard 2 100X Biotinylated CYP7A1 Detector Antibody 1 x 120 uL 100X Biotinylated CYP7A1 Detector Antibody 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Cytochrome P450 7A1 (CYP7A1) . No significant cross-reactivity or interference between Cytochrome P450 7A1 (CYP7A1) and analogues was observed.