The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration. Predicted molecular weight: 56 kDa.
Use a concentration of 5 µg/ml.
RelevanceThe Cytochrome P450 superfamily of enzymes is one of three enzyme systems which metabolize the fatty acid arachadonic acid (AA) to regulators of vascular tone. P450 enzymes are monooxygenase enzymes which require several co-factors such as nicotinamide adenine dinucleotide phosphate (NADPH) and P450 reductase. There are over 200 known genes which encode P450s. Epoxygenases are those P450s which metabolize AA to epoxyeicosatrienoic acid (EETs) and omega-hydroxylases are those P450s which produce 19- and 20-hydroxyeicosatetraenoic acids (19- and 20-HETE). As well as fatty acid metabolism, P450s also metabolize many drugs and toxins. Cytochrome P450 3A4 is abundantly expressed in liver and small intestine and is inducible by barbiturates, glucocorticoids and rifampicin.
Cytochrome P450 2B1 + 2B2 was immunoprecipitated using 0.5mg Mouse Liver tissue, 5µg of Mouse monoclonal to Cytochrome P450 2B1 + 2B2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver tissue lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab22719.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.