Volume 37, Issue 6 (November 1992)
The Application of Immunoblotting to the Phenotyping of Haptoglobin
An immunoblotting method for phenotyping haptoglobin in serum and blood-stains has been developed. Haptoglobin isoproteins were separated by polyacrylamide gradient gel electrophoresis and then transferred to nitrocellulose by electroblotting. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm thick gels. Nitrocellulose blots were developed by double antibody enzyme immunoassay. The detection limit for serum and bloodstains was improved 16 times compared to conventional staining using O-tolidine. The method could detect haptoglobin phenotypes from 0.001 μl of whole blood. This detection limit is approximately 8 times lower than that of group specific-component analysis by immunoblotting.