During electrophoresis generation of heat is the major problem which leads to

ncreased rate of diffusion of sample and buffer ions.

Thermal instability which include denaturation of proteins.

A decrease of buffer viscosity.

Convection current leads mixing of separated samples

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#Unit 13. Methods in Biology

He received Nobel Prize in 1948 for developing technique for electrophoretic separation of substances.

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#Unit 13. Methods in Biology

Overall, there are three pillars electrophoresis

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#Unit 13. Methods in Biology

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

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#Unit 13. Methods in Biology

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

Slab gels can develop a “Smiling Effect” When in a gel more Joule heat is produced than can be dissipated, leads to the higher mobility of the ions in the center, resulting in an uneven distribution of the zones, These zones bend up at the two lateral sides, causing “smiling.” 

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#Unit 13. Methods in Biology

Phenomenon of Smiling Effect is largely applicable in Gel Electrophoresis &  PAGE