Sensitivity, ease of use, and shelf life are all considerations in choosing detection systems for Western blots and dot blots. Chemiluminescent substrates allow the sensitive detection of targeted proteins in immunoblotting protocols with stable, hard-copy results on film-all without the hazards and other problems associated with the use of radioactive probes.
Numerous chemiluminescent substrates are commercially available. They generally consist of luminol and H2O2 with undisclosed chemical enhancers and produce luminescent reactions with horseradish peroxidase.
Comparison of SuperSignal and SuperSignal ULTRA: Seven gels, each containing duplicates of 5, 10 and 20 microgram samples of beta-actin were blotted, incubated with 1:100,000 dilutions of primary and secondary antibody for 30 minutes and exposed either to SuperSignal or SuperSignal Ultra for five minutes. Densitometric values were averaged for each beta-actin concentration. Data are summarized in the bar graph, which was adapted from D.L. Mattson and T.G. Bellhumeur, Analytical Biochem. 240: 306-308, 1996,...
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