Rat serum EU143 was raised against the predicted C-terminal 353 residues of Ce-titin. Antigens was expressed as GST fusion proteins in E. coli. cDNAs encoding antigenic polypeptides were amplified by RT-PCR using Pfu polymerase (Stratagene) and primers containing added XhoI and EcoRI sites. The primers for EU143 were 5'-GTACGAATTCAAATTCATCTCAGTGACACCAGG-3' and 5'-GATGCTCGAGCTACCGACGAATCGTTCTTCGTTG-3', respectively. Amilified product was cloned into pGEX-6P-1 (Amersham Pharmacia Biotech).T he GST fusion proteins were expressed and purified. Milligram quantities of fusion proteins were supplied to Spring Valley Laboratories (Sykesville, MD) for antiserum production.
ttn-1