An enzyme-linked immunoadsorbent assay for measuring cytochrome b5 and NADPH-cytochrome P-450 reductase in rat liver microsomal fractions. Evidence for functionally inactive protein.
Immunoreactive cytochrome b5 and NADPH-cytochrome P-450 reductase (EC 1.6.2.4) from rat liver microsomal fractions were measured by using an enzyme-linked immunoadsorbent assay (e.l.i.s.a.) as a function of age, sex and type of inducer (phenobarbital or 3-methylcholanthrene), and the values were compared with those obtained by spectral measurement (for cytochrome b5) or enzymic assay (for reductase). In untreated animals, there was more cytochrome b5 and NADPH-cytochrome P-450 reductase when measured by an e.l.i.s.a. than was seen spectrally or enzymically. However, for microsomal preparations from phenobarbital-pretreated animals, spectrally obtained values for cytochrome b5 and immunoreactive-cytochrome b5 values were similar. Values from control animals suggest that there is about 20-30% more immunoreactive cytochrome b5 than that which is spectrally detectable.
D000367:Age Factors; D000818:Animals; D003573:Cytochrome b Group / Q000032:analysis*; D015786:Cytochromes b5; D004797:Enzyme-Linked Immunosorbent Assay; D066298:In Vitro Techniques; D008748:Methylcholanthrene / Q000494:pharmacology; D008862:Microsomes, Liver / Q000032:analysis* / Q000187:drug effects / Q000201:enzymology; D009251:NADPH-Ferrihemoprotein Reductase / Q000032:analysis*; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012737:Sex Factors; D013053:Spectrophotometry
D003573:Cytochrome b Group; D008748:Methylcholanthrene; D015786:Cytochromes b5; D009251:NADPH-Ferrihemoprotein Reductase; D010634:Phenobarbital