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Immunoblotting with monoclonal antibodies: importance of the
blocking solution
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
ID 153430
Author(s) Hauri, H.P.; Bucher, K.
Author(s) at UniBasel Hauri, Hans-Peter;
Year 1986
Title Immunoblotting with monoclonal antibodies: importance of the blocking solution
Journal Analytical Biochemistry
Volume 159
Number 2
Pages / Article-Number 386-389
Keywords *Antibodies; Monoclonal; Epitopes; Humans; *Immunologic Techniques; Intestinal
Mucosa/enzymology; Solutions; Sucrase-Isomaltase Complex/immunology
Four commonly used blocking agents, i.e., fetal calf serum, mammalian gelatin-Nonidet-P40, fish gelatinNonidet-P40, and defatted powdered milk were compared with respect to their efficiency to block the
nonspecific background and to promote maximal immunoreactivity of monoclonal antibodies against human
intestinal sucrase-isomaltase during immunoblotting. Two of five monoclonal antibodies were found to react
with the electroblotted enzyme. However, one of the reacting antibodies gave optimal results with fish gelatinNonidet-P40 and the other with defatted powdered milk, while fetal calf serum lead to unacceptably high
backgrounds. The results suggest that some of the difficulties encountered with monoclonal antibodies in
immunoblotting may be due to inappropriate blocking conditions.
ISSN/ISBN 0003-2697
Full Text on edoc
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/2435189