Immunoblotting optimization of proteins separated by two dimensional electro-phoresis: a proteomics approach for determining new autoantigens in autoimmune hepatitis type 1

Autoimmune Hepatite type 1 (AIH-1) is serologically diagnosed by presence of anti nuclear antibody and/or anti smooth muscle antibody which molecular target of them is not known. A proteomics approach based on two dimensional electrophoresis (2DE) and immunoblotting was utilized in order to search out possible profiles of a hepatoma cell line, HepG2, were established by 2DE and blotted to the nitrocellulose membrane.We showed the crucial criteria in selecting the significant immunostained spots following immunoblotting of HepG2 protein profile by patientsُ sera mixture and healthy control counterparts.

Keywords:

Autoimmune hepatitis , two dimensional electrophoresis , immunoblotting , proteomics