Characterization of a synthetic peptide antibody recognizing rat kidney renin and prorenin.
A fourteen amino acid peptide from the cDNA predicted rat renin amino acid sequence was synthesized and used to produce a polyclonal antibody. The ability of this antibody to recognize rat renin was characterized and compared to that of a previously tested polyclonal anti-rat renin antibody raised against purified rat renin to verify its usefulness as a reagent in renin processing. The anti-renin peptide antibody and anti-rat renin antibody were evaluated for specificity of recognition of rat renin by immunohistochemical staining and immunoblotting. The ability of the anti-renin peptide antibody to inhibit renin enzymatic activity and to immunoprecipitate rat recombinant protein was also tested. Anti-renin peptide antibody and anti-rat renin antibody had identical recognition of rat renin and prorenin in immunohistochemical stains of rat kidney section and immunoblots. However the anti-renin peptide antibody does not inhibit renin enzymatic activity nor does it immunoprecipitate recombinant rat prorenin. We have produced and characterized an antirenin peptide antibody which recognizes denaturated rat renin and prorenin in a way that is identical to that of an antibody raised against native rat renin. The anti-renin peptide antibody does not recognize nondenatured renin because it did not inhibit renin enzymatic activity nor was it able to immunoprecipitate renin or prorenin. The technique of using synthetic peptides to produce antibody recognizing at renin is useful because low yields of native renin purified from rat kidney are generally insufficient for antibody production. This limits the widespread availability of antibodies that recognize rat renin.(ABSTRACT TRUNCATED AT 250 WORDS)
Norling
L L
LL
Pediatric Nephrology Unit, Massachusetts General Hospital, Boston, USA.
Gomez
R A
RA
Inagami
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D004792:Enzyme Precursors / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D007668:Kidney / Q000201:enzymology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D051381:Rats; D011921:Rats, Inbred WKY; D011994:Recombinant Proteins / Q000276:immunology; D012083:Renin / Q000276:immunology*
D004792:Enzyme Precursors; D000939:Epitopes; D010455:Peptides; D011994:Recombinant Proteins; D012083:Renin