title: Mechanism of Coomassie Brilliant Blue G-250 binding to cetyltrimethylammonium bromide: an interference with the Bradford assay.
abstract: The Bradford protein assay is a popular method because of its rapidity, sensitivity, and relative specificity. This method is subject to some interference by nonprotein compounds. In this study, we describe the interference of cetyltrimethylammonium bromide (CTAB) with the Bradford assay. This interference is based on the interaction of Coomassie Brilliant Blue G-250 (CBB) with this cationic detergent. This study suggests that both electrostatic and hydrophobic interactions are involved in the interaction of CTAB and CBB. The anionic and neutral forms of CBB bind to CTAB by electrostatic attraction, which accelerates hydrophobic interactions of these CBB forms and the hydrophobic tail of CTAB. Consequently, the hydrophobic regions of the dominant free cationic form of CBB dye compete for the tail of CTAB with two other forms of the dye and gradually displace the primary hydrophobic interactions and rearrange the primary CBB-CTAB complex. This interaction of CTAB and CBB dye produces a primary 650-nm-absorbing complex that then gradually rearranges to a complex that shows an absorbance shoulder at 800-950 nm. This study conclusively shows a strong response of CBB to CTAB that causes a time-dependent and nearly additive interference with the Bradford assay. This study also may promote an application of CBB for CTAB quantification.
authors.affiliation: Department of Clinical Biochemistry, School of Medicine, Tehran, Iran. amminian@yahoo.com
authors.forename: Mahdi
authors.initials: M
authors.lastname: Aminian
authors.forename: Fariba
authors.initials: F
authors.lastname: Nabatchian
authors.forename: Asad
authors.initials: A
authors.lastname: Vaisi-Raygani
authors.forename: Mojgan
authors.initials: M
authors.lastname: Torabi
mesh_terms: D016477:Artifacts; D001681:Biological Assay / Q000592:standards*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D015394:Molecular Structure; D054730:Protein Interaction Domains and Motifs; D012394:Rosaniline Dyes / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry
chemical_list: D002593:Cetrimonium Compounds; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine; C004692:coomassie Brilliant Blue; D000077286:Cetrimonium