Overall, there are three pillars electrophoresis

Current / voltage

Medium - (buffer/liquid) - with a precise pH value and a constant ionic strength.

 Analytes - A compound, which is to be detected or assayed in a given sample.

Electrophoretic resolution mainly can be change by manipulating current and buffer system.

The higher the buffer concentration, the more electric power is applied, resulting in higher heat (joule effect) development.

Smiling Effect, Venetian Blind Effect & Electroendosmosis reduces the efficiency of electrophoresis.

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#Unit 13. Methods in Biology

SDS (H3C-(CH2 )10 –CH2 -OSO3 – Na+ ) is an anionic detergent used to create negative on sample  On average, one SDS molecule binds for every two amino-acid residues.

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#Unit 13. Methods in Biology

SDS–PAGE are firstly boiled for 5 min in sample buffer containing - mercaptoethanol or dithiothreitol ( DTT) and SDS. Mercaptoethanol or DTT reduce any disulfide bridges

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#Unit 13. Methods in Biology

In conventional discontinuous gel electrophoresis two type of GEL system is used on top with comb there is a stacking gel with pH 6.8 and below resolving with gel pH 8.8 phenomenon known as isotachophoresis.

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#Unit 13. Methods in Biology

The band-sharpening effect relies on the fact that negatively charged glycinate ions (in the electrophoresis buffer) have a lower electrophoretic mobility than do the protein–SDS complexes, which, in turn, have lower mobility than the chloride ions (Cl − ) of the loading buffer and the stacking gel buffer. 

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#Unit 13. Methods in Biology

More recently Different from conventional gels, the matrix of single gel system introduced  which contains  three amino acids (serine, glycine and asparagine).

#Id: 2495

#Unit 13. Methods in Biology

The relationship between acrylamide gel concentration and protein fractionation range