NCJ Number
139773
Journal
Journal of Forensic Sciences Volume: 37 Issue: 6 Dated: (November 1992) Pages: 1652-1655
Date Published
1992
Length
4 pages
Annotation
This article describes the method and results of using an immunoblotting method for phenotyping haptoglobin in serum and blood stains.
Abstract
Blood stains were prepared by soaking cotton cloth with fresh whole blood collected from laboratory staff. Portions of stained cloth were extracted in 20-percent sucrose in tank buffer. Whole blood and serum samples were diluted with 20-percent sucrose/tank buffer. Separation of haptoglobin isoproteins was achieved by vertical gel electrophoresis in polyacrylamide gradient gels. The transfer of haptoglobin proteins from unstained gels to nitrocellulose was achieved by electroblotting at 70 volts for 3 hours, using an LKB 2051 Midgit Multiblot Electrotransfer unit. Nitrocellulose blots were developed by a modification of the method previously described. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm-thick gels. The detection limit for serum and blood stains was improved 16 times compared to conventional staining using O- toluidine. The method could detect haptoglobin phenotypes from 0.001 ml of whole blood. This detection limit is approximately eight times lower than that of group-specific component analysis by immunoblotting. 2 figures and 5 references