Premium
Detection of proteins after immunoblotting on nitrocellulose using fluorescent antibodies
Author(s) -
Abuharfeil Nizar M.,
Atmeh Ragheb F.,
AboShehada Mahmoud N.,
ElSukhon Saeb N.
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150120917
Subject(s) - nitrocellulose , antibody , chemistry , immunoglobulin light chain , conjugated system , fluorescence , chromatography , microbiology and biotechnology , enzyme , biochemistry , monoclonal antibody , primary and secondary antibodies , biology , immunology , physics , organic chemistry , quantum mechanics , membrane , polymer
A Western blot method is described utilizing 5‐dimethylaminonaphthalene‐1‐sulfonyl (dansyl chloride)‐conjugated antibodies to detect proteins transferred onto nitrocellulose. Four proteins of different molecular weight, including human albumin, gamma heavy chain, kappa and lambda light chains of immunoglobulins, were tested. Labeling with the fluorochrome was compared with the method of enzyme‐conjugated antibodies and was found to have similar sensitivity, specificity, and safety. Moreover, this method is less expensive, and the labeled antibodies can be prepared fresh in a short time and are free from the disadvantages of enzyme label whose activity is affected by the presence of activators or inhibitors in the tested specimens.