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SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) is a discontinuous electrophoretic system developed by Ulrich K. Laemmli which is commonly used as a method to separate proteins with molecular masses between 5 and 250 kDa.
Picture of an SDS-PAGE. The molecular markers (ladder) are in the left lane. Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility.
Proteins separated by SDS-PAGE, Coomassie brilliant blue staining. Protein electrophoresis is a method for analysing the proteins in a fluid or an extract. The electrophoresis may be performed with a small volume of sample in a number of alternative ways with or without a supporting medium, namely agarose or polyacrylamide.
Separate just by mass is commonly achieved using SDS-PAGE. SDS denatures the proteins, breaks apart most complexes, and approximately equalizes the mass-to-charge ratios. SDS must be done as the second, perpendicular dimension, as it breaks apart complexes (rendering native PAGE impossible) and equalizes mass-to-charge ratios (rendering IEF ...
The combination of Na[BH 4] with carboxylic acids results in the formation of acyloxyborohydride species other than sodium triacetoxyborohydride. These modified species can perform a variety of reductions not normally associated with borohydride chemistry, such as alcohols to hydrocarbons and nitriles to primary amines.
Reductive amination, sometimes called the Borch reaction, is the conversion of a carbonyl into an amine through an intermediate imine. [4] The carbonyl is first treated with ammonia to promote imine formation by nucleophilic attack. The imine is then reduced to an amine by sodium cyanoborohydride. This reaction works on both aldehydes and ketones.
Isoelectric focusing is the first step in two-dimensional gel electrophoresis, in which proteins are first separated by their pI value and then further separated by molecular weight through SDS-PAGE. Isoelectric focusing, on the other hand, is the only step in preparative native PAGE at constant pH. [5]
Following 2D SDS PAGE the proteins can be transferred to a polyvinylidene difluoride (PVDF) blotting membrane for further analysis. Edman degradations can be performed directly from a PVDF membrane. N-terminal residue sequencing resulting in five to ten amino acid may be sufficient to identify a Protein of Interest (POI). [citation needed]