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Preparation of mono specific antibodies against 2 forms of rat liver cytochrome p 450 and quantitation of these antigens in microsomes


, : Preparation of mono specific antibodies against 2 forms of rat liver cytochrome p 450 and quantitation of these antigens in microsomes. Archives of Biochemistry & Biophysics 192(2): 524-532

Antibodies prepared against purified cytochrome P-450 and P-448 from phenobarbital- and 3-methylcholanthrene-pretreated rats recognize several forms of hepatic cytochrome P-450. These antibodies were made monospecific for a single form of cytochrome P-450 by immunoadsorption with partially purified solid-phase cytochrome P-450 from rats treated with a different inducer than that used for isolation of the antigen. Each monospecific antibody did not react with different forms of cytochrome P-450 present in the heterologous antigen preparation. These monospecific antibodies, covalently bound to Sepharose, were used to purify the antigens (catalytically inactive) from microsomes in a single step. The high specificity of these antibodies for a single form of cytochrome P-450 was used to quantitate 2 forms of cytochrome P-450 in rat liver microsomes by radial immunodiffusion. The percentage of the total cytochrome P-450 in microsomes that is represented by each of these 2 forms of cytochrome P-450 varied from 3-89% depending on the xenobiotic pretreatment of the rats.

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