Interference of the detergent Tween 80 in protein assays.
The nonionic detergent Tween 80, which has been widely used to stimulate protein secretion in bacterial and fungal systems, caused interferences in three protein determination methods. The OD595 developed in the Coomassie blue dye-binding assay with a variety of purified proteins in the presence of Tween 80 was 1.6 to 3.4 times greater than that observed without detergent. These differences could not be attributed totally to the rapid color development in the assay with Tween 80 alone. Crude concentrated extracellular bacterial proteins shaken overnight with Tween 80 yielded an altered fractionation pattern on size exclusion chromatography and 10-fold increased color with an absorption spectrum in the dye-binding assay different from that of bacterial proteins shaken without detergent. In the bicinchoninic acid method, the detergent caused a 2- to 3-fold increase in OD562 due largely to contaminating peroxides which could be removed by treatment with catalase. In the Folin phenol method, the detergent caused a slight precipitate, but residual interference was not detectable in filtered assay mixtures.
Stutzenberger
F J
FJ
Department of Microbiology, Clemson University, South Carolina 29634.
D000192:Actinomycetales / Q000737:chemistry; D016477:Artifacts; D001426:Bacterial Proteins / Q000032:analysis; D004798:Enzymes / Q000032:analysis; D007202:Indicators and Reagents; D011136:Polysorbates; D011506:Proteins / Q000032:analysis*; D013053:Spectrophotometry / Q000379:methods
D001426:Bacterial Proteins; D004798:Enzymes; D007202:Indicators and Reagents; D011136:Polysorbates; D011506:Proteins