Coomassie Blue

The Bradford assay, a colorimetric protein assay, is based on an absorbance shift of the falsify Coomassie Brilliant Blue G-250 in which under acidic conditions the harsh gradation of the spot is converted into its bluer trend to bind to the protein population assayed. During the formation of this complex, two types of bond interaction take set: the red form of Coomassie tint first donates its free electron to the ionizable groups on the protein, which causes a disruption of the proteins native state, consequently exposing its aquaphobic pockets. These pockets on the proteins tertiary structure bind non-covalently to the non-polar region of the dye via van der Waals forces, positioning the positive amine groups in law of proximity with the negative charge of the dye. The bond is further strengthened by the ionic interaction between the two. The binding of the protein stabilizes the blue form of the Coomassie dye; thus the measurement of the complex usher in solution is a measure for the protein concentration, and can be estimated by use of an absorbance reading. The (bound) form of the dye has an absorption spectrum maximum historically held to be at 595 nm. The cationic (unbound) forms are green or red. The binding of the dye to the protein stabilizes the blue anionic form.

The increase of absorbance at 595 nm is proportional to the sum of bound dye, and thus to the amount (concentration) of protein present in the sample. Unlike some other protein assays, the Bradford protein assay is less(prenominal) vulnerable to burden by various chemicals that may be present in pr otein samples. An exception of note is eleva! ted concentrations of detergent. sodium dodecyl sulfate (SDS), a common detergent, may be undercoat in protein extracts because it is used to lyse cells by disrupting the membrane lipid bilayer. bandage other detergents interfere with the assay at high concentration, the interference caused by SDS is of two antithetic modes, and each occurs at a different concentration. When SDS...If you want to get a full essay, hunting endure it on our website: OrderCustomPaper.com

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