The use of nitrocellulose blotting for the study of hepatitis B surface antigen electrophoresed in agarose gels.
Nitrocellulose-protein blotting of serum electrophoresed in agarose gels has been adapted for the study of hepatitis B surface antigen (HBsAg). 125I-labeled anti-HBs was used as the antigen probe, and the electrophoretic migration was monitored by autoradiography. The method required 3 microliter or less of serum and could detect as little as 1 pg of purified HBsAg. Typically, we observed two bands of HbsAg; a moving band which migrated about one-third the distance moved by human serum albumin and a non-migratory band which remained at the loading site. Some examples of the use of the method include: (1) empirical methods for correlating HBsAg concentration in serum to film darkness; (2) observations of mobility changes in serial sera from dialysis patients with chronic HBsAg antigenemia; and (3) detection of related antigens such as antigen from the PLC/PRF/5 hepatoma tissue culture line and the cross-reacting woodchuck patients hepatitis virus surface antigen (WHsAg).
D000818:Animals; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D003101:Collodion / Q000494:pharmacology*; D004587:Electrophoresis, Agar Gel; D006509:Hepatitis B / Q000175:diagnosis; D006514:Hepatitis B Surface Antigens; D006801:Humans; D007158:Immunologic Techniques; D008137:Longitudinal Studies; D008392:Marmota; D009038:Motion; D010209:Paper; D011489:Protein Denaturation
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D006514:Hepatitis B Surface Antigens; D003101:Collodion