Sodium dodecyl sulphate (SDS) is an anionic detergent which denatures proteins by "wrapping around" the polypeptide backbone - and SDS binds to proteins fairly specifically in a mass ratio of 1.4:1. In so doing, SDS confers a negative charge to the polypeptide in proportion to its length - ie: the denatured polypeptides become "rods" of negative charge cloud with equal charge or charge densities per unit length. It is usually necessary to reduce disulphide bridges in proteins before they adopt the random-coil configuration necessary for separation by size: this is done with 2- mercaptoethanol or dithiothreitol. In denaturing SDS-PAGE separations therefore, migration is determined not by intrinsic electrical charge of the polypeptide, but by molecular weight
And for the actual experiment beyond the denaturing: PAGE stands for polyacylamide gel electrophoresis. This is a procedure that separates proteins by size by running them through a gel "matrix" so that the smaller ones travel faster that the larger ones. This is achieved by creating an electric field with the sds-protein complex traveling toward the positively charged end of the gel. Once the smallest proteins have traveled most of the way across the gel the current is turned of and the gel is removed and stained with dye that binds protein so that you can see where it is in the gel.
Due to many proline residues it migrates slower on sds page and appears heavier than it is.
It is the gel of choice for SDS PAGE
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SDS-PAGE AGAROSE CAPILLARY SEQUENCING TO NAME A FEW
SDS - PAGE is apparently used to seperate proteins. The proteins are by nature different sizes. SDS works as a stabilizer by separating proteins according to similar forms.
may be because of toomany disulfide linkages
CTAB is a cationic detergent. It is generally considered to be less denaturing than SDS. It is typically used when it is desired to maintain enzymatic activity. It is also typically used in protein refolding.
glycine molecular weight high so mobility also high so using in SDS PAGE
Due to many proline residues it migrates slower on sds page and appears heavier than it is.
The major drawback is that treatment with SDS denatures the protein, meaning you are not looking at it in its natural state.
It is the gel of choice for SDS PAGE
Laemmli U. K.
SDS-PAGE method
Glycine increases the mobility of the gel.
Electrophoresis is the method that could be used to further separate two bands from the same protein fraction after SDS-PAGE.
break the S-S bonds in a protein
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