The Preparation Of AKO62675 Antibody And Expression Research In The Bladder Carcinoma

Objective: 1.Bioinformatics analysis the full length sequence of cDNA which had been obtained in the earlier stage,submit it to the NCBI and get GenBank accession number:KR780251,protein ID:AKO62675.2.Choose the most differences between AKO62675 and CK20 to design a polypeptide,and prepare antibody: Anti-KR.3.According to the expression of AKO62675 in the bladder carcinoma tissue,glandular cystitis tissue and normal bladder mucosa membrane tissue,analyze whether AKO62675 contribute to the occur of bladder carcinoma.Methods: 1.To explore chromosome localization and protein analysis of the full length sequence of cDNA,submit it to the NCBI and get Gen Bank accession number: KR780251,protein ID:AKO62675 by sequin software.2.Choose the most differences between AKO62675 and CK20 to design a polypeptide,then prepare antibody named Anti-KR by immunity to rabbit.3.Detect AKO62675 expression in 40 cases of bladder carcinoma tissue,15 cases of glandular cystitis tissue and 23 cases of normal bladder mucosa membrane tissue by immunohistochemical,compare the AKO62675 expression in these different tissues,analyze if AKO62675 contribute to the occur and development of bladder carcinoma.Results: 1.We found the sequence had a different localization in the chromosome compare to CK20,submit it to the NCBI and get GenBank accession number:KR780251,protein ID:AKO62675.2.Detect the polyclonal antibody named Anti-KR by ELISA,the titer beyond 1:64000 and reached the measurement demand.3.Immunohistochemical staining of bladder carcinoma tissue,glandular cystitis tissue and normal bladder mucosa membrane tissue: the positive rate is 34.78% in normal bladder mucosa membrane tissue,the positive rate is 100% in bladder carcinoma tissue and glandular cystitis tissue,while the dyeing is more shallow in glandular cystitis tissue than carcinoma tissue.Conclusions: 1.The full length of EST fragment KR780251 has 1839 bp and contains complete ORF.The protein of AKO62675 encoded by KR780251 may be the mutant gene of normal bladder tissue.2.AKO62675 has high expressiveness in bladder carcinoma.