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Since your protein is relatively big, approx. 10% or lower fixed percentage acrylamide gels would do the job. In the case of other targets along with 88kda protein as Adam B Shapiro indicated,...
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Since your protein is relatively big, approx. 10% or lower...
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Dependent on the molecular weight of the protein(s) of interest different formulations of acrylamide gel are required to gain optimal separation of proteins. Gels can either use a single concentration of acrylamide where they are optimised for a narrow range of molecular weights (see section 1) or be gradients of acrylamide concentration where ...
smaller pore size resolving gel begins to separate the proteins based on molecular weight only, since the charge-to-mass ratio is equal in all the proteins of the sample; F, The individual proteins are separated into band patterns ordered according to their molecular weights.
Typical gel compositions are between 7.5 and 20% for single-percentage gels, and typical gradients are 4–15% and 10–20%. Use protein migration charts and tables to select the gel type that offers optimum resolution of your sample (see figure below).
The porosity of the gel is determined by the relative concentration of acrylamide to cross-linker and by the total percentage of monomers. The final concentration of acrylamide depends on the sample under study with high acrylamide concentrations giving better resolution of low molecular mass proteins, and vice versa.
Polyacrylamide gels can be considered as a filter with pore size that can be modulated by changing acrylamide percentage. Higher percentage will generate smaller pore size and will preferentially be used to separate protein with low molecular weight.
15 de abr. de 2014 · Two dimensional polyacrylamide gel electrophoresis (2-DE) is considered a powerful tool used for separation and fractionation of complex protein mixtures from tissues, cells, or other biological samples. It allows separation of hundreds to thousands of proteins in one gel.
