A blocking agent and a blocking step are not needed in ELISA, immunostaining dot-blots and western blots.
The effect of blocking and non-blocking was compared in ELISA, immunostaining dot-blots and western blots using phosphate-buffered saline with and without the addition of Tween 20 for washes and dilutions. The results indicated that when Tween 20 was included in phosphate-buffered saline, a blocking agent and a blocking step were not required in immunoassays. Moreover, the use of protein-based blocking agents was found to somewhat lower the reactivity between the antibody and the antigen, possibly due to steric hindrance.
Mohammad
K
K
Department of Biology, Virginia Polytechnic Institute, Blacksburg 24061.
Esen
A
A
D000818:Animals; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D015153:Blotting, Western / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera; D015151:Immunoblotting / Q000379:methods; D008892:Milk; D011136:Polysorbates; D012710:Serum Albumin, Bovine; D013194:Staining and Labeling / Q000379:methods; D015028:Zein / Q000276:immunology
D007106:Immune Sera; D011136:Polysorbates; D012710:Serum Albumin, Bovine; D015028:Zein