Phosphorylation of T14 and Y15 in this G-loop

introduces negative charge that electrostatically interferes with the binding of ATP in the catalytic pocket.

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#Unit 13. Methods in Biology

Gradient Gels electrophoresis are performed using gradient of  acrylamide Concentration that varies uniformly from, typically, 5% at the top of the gel to 25% acrylamide at the bottom of the gel.

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#Unit 13. Methods in Biology

There are two major advantages to running gradient gels. 

First, a much greater range of protein Mr values can be separated than on a fixed- percentage gel.

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#Unit 13. Methods in Biology

The second advantage of gradient gels is that proteins with very similar M r values may be resolved.

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#Unit 13. Methods in Biology

The higher no. of band will be observed in gradient gel electrophoresis comparison to uniform gel SDS and Native gel electrophoresis.

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#Unit 13. Methods in Biology

Isoelectric Focusing Gels is based on the separation of molecules according to their different isoelectric points at which the show zero electrophoretic mobility.

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#Unit 13. Methods in Biology

In the Isoelectric Focusing pH gradient is formed by the introduction into the gel of compounds known as ampholytes, which are complex mixtures of synthetic polyamino-polycarboxylic acids Ampholytes can be purchased in different pH ranges covering a wide band (e.g. pH 3–10).