SDS PAGE Sample Buffer (5X) (Non-Reducing) w/o DTT w/ BPB

It is the most commonly used sample buffer for SDS-PAGE of non-denatured proteins It is a ready-to-use 5X solution without dithiothreitol (DTT) and Beta-mercaptoethanol (BME) as a reducing agent. This buffer can be used as a loading buffer in all kinds of denaturing gels and are compatible with Coomassie, silver staining and Western blotting. The 5X concentration of this buffer facilitates the loading of larger sample volume per well compared to the traditional 2X loading buffer.Role of reagents: Tris-HCl: as a buffering substance. The pH play an essential role in preserving peptide bonds from breaking apart. SDS: Proteins comes in different sizes and charges. SDS is an anionic detergent that helps in linearizing (by denaturing) the proteins and bringing a net negative charge to the proteins irrespective of the initial charge. Glycerol: The high density (thickening of the solution) of glycerol ensures the sample moves down into the well. Bromophenol Blue: visually indicates the location (tracking dye) of the sample in the gel.

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