ISSN:
0173-0835
Keywords:
Chemistry
;
Biochemistry and Biotechnology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
Notes:
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the immunoblotting technique, were applied to structural analysis of a single protein (alpha-2-macroglobulin, α-2-M) in human serum or plasma without prior purification of this protein. The use of immunoglobulin fractions of four monospecific polyclonal antibody preparations to α-2-M in the immunoblotting step showed marked unspecific binding to the proteins fixed onto nitrocellulose. Immunospecific purification of the antibodies by two different methods, and the introduction of biotinylated antibodies and avidin-peroxidase, removed the background binding. When reduced proteins were fixed onto nitrocellulose a quantitative loss of α-2-M antigen reactivity was observed, compared to identical nonreduced samples. The degradation pattern of α-2-M in serum or plasma corresponded to the pattern observed with purified α-2-M. Thus, the described immunoblotting technique, applied directly to serum or plasma, could detect all not heat-denaturated α-2-M products as seen on Coomassie-stained polyacrylamide gels when analyzing purified α-2-M.
Additional Material:
4 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elps.1150060506
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