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Gel Electrophoresis of Proteins Articles

Isoelectric Focusing

By National Diagnostics | September 9, 2011 | Comments Off on Isoelectric Focusing

Conventional electrophoresis techniques can separate up to 100 different proteins on one run. Typically, cell or tissue extracts contain thousands of proteins, most of which will not be resolved into single bands using a separation…

Immuno-Electrophoresis / Immuno-Diffusion

By National Diagnostics | September 9, 2011 | Comments Off on Immuno-Electrophoresis / Immuno-Diffusion

Antibodies are produced by the immune system in response to foreign macromolecules. Each antibody binds specifically to one feature (epitope) on one macromolecule (antigen). This allows the use of antibodies for the detection and quantitation…

Activity Stains

By National Diagnostics | September 9, 2011 | Comments Off on Activity Stains

Samples to be run on native gels should be prepared in a way that minimizes the denaturation of the proteins. Avoid heat, strong detergents, foaming, and over-dilution. In addition, the activity of endogenous proteases must…

Gel Preparation for Native Protein Electrophoresis

By National Diagnostics | September 9, 2011 | Comments Off on Gel Preparation for Native Protein Electrophoresis

The basic protocols for preparing Native PAGE gels are the same as for discontinuous SDS PAGE gels, substituting non-SDS buffers for those containing SDS, as follows: Casting Native Protein Gels Prepare resolving gel and stacking…

Sample Preparation for Native Protein Electrophoresis

By National Diagnostics | September 9, 2011 | Comments Off on Sample Preparation for Native Protein Electrophoresis

Samples to be run on native gels should be prepared in a way which minimizes denaturation of the proteins. Avoid heat, strong detergents, foaming and over-dilution. In addition, the activity of endogenous proteases must be…

Native Protein Electrophoresis

By National Diagnostics | September 9, 2011 | Comments Off on Native Protein Electrophoresis

Proteins run on PAGE in the absence of SDS will separate on the basis of their charge to mass ratio. While native (nondenaturing) PAGE does not provide direct measurement of molecular weight, the technique can…

Peptide Mapping

By National Diagnostics | September 7, 2011 | Comments Off on Peptide Mapping

Peptide mapping involves controlled cleavage of a pure protein with small amounts of a pure protease to generate peptides of characteristic, reproducible sizes. These peptides can be separated on PAGE to produce a “fingerprint” characteristic…

Measuring Molecular Weight with SDS-PAGE

By National Diagnostics | September 7, 2011 | Comments Off on Measuring Molecular Weight with SDS-PAGE

The mobility (Rf) of a molecule in gel electrophoresis is determined by its free solution mobility, Y0 (= mobility in a gel of zero %) and the sieving action of the gel matrix. In denaturing…

Casting Gradient Gels

By National Diagnostics | September 7, 2011 | Comments Off on Casting Gradient Gels

Gradient gels are cast with a higher concentration of acrylamide at the bottom than the top. Gradient gel applications include the determination of protein molecular weights and the separation of molecules which co-migrate on uniform…

Sample Preparation for SDS-PAGE

By National Diagnostics | September 6, 2011 | Comments Off on Sample Preparation for SDS-PAGE

SDS is a powerful detergent, which will solubilize many cells and tissues. This greatly facilitates sample preparation for SDS PAGE because most samples will be completely dissolved by heating to 95°C in loading buffer (detailed…