Synthetic peptide antigens elicit monoclonal and polyclonal antibodies to cytochrome P450 IA2.
Two peptide sequences from cytochrome P450 IA2 were synthesized, coupled to ovalbumin and used as antigens to generate anti-peptide monoclonal and polyclonal antibodies. Antisera to both peptides reacted with rat IA2 but not the structurally similar IA1 form as determined by enzyme-linked immunosorbent assay. However, antisera to both peptides detected both rat IA2 and IA1 on immunoblots. In addition immunoblots of human liver microsomes revealed that both antisera recognized human IA2, but not IA1. Monoclonal antibodies generated against one of the peptides recognized rat IA2 and IA1 but did not detect human IA2. These results demonstrate the utility of anti-peptide antisera as a practical approach for the generation of P450 specific antibodies.
Myers
M J
MJ
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, MD 20892.
Liu
G
G
Miller
H
H
Gelboin
H V
HV
Robinson
R C
RC
Friedman
F K
FK
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D003577:Cytochrome P-450 Enzyme System / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D007527:Isoenzymes / Q000276:immunology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D007527:Isoenzymes; D010446:Peptide Fragments; D003577:Cytochrome P-450 Enzyme System