Synthetic peptide antisera: their production and use in the cloning of matrix proteins

Small amounts (0.1-1.0 nmole) of purified (greater than 90%) proteins isolated from small quantities of connective tissues can readily be microsequenced and 20 to 40 N-terminal amino acids determined. A synthetic peptide (10 or more amino acids long) can be produced and either injected directly into rabbits for antiserum production (generally peptides greater than 20 amino acids in length) or conjugated to a carrier protein prior to injection. The antisera have proven useful in the isolation of cDNA clones, for both locating clones producing the protein in expression libraries and in the subsequent confirmation of the sequence (preferably using a part of the sequence not directly used in the production of the original antiserum).