User:Laura Flynn/Notebook/Experimental Biological Chemistry/2011/10/25

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Objective

The purpose of today's procedure is to run a protein gel using a Bio Rad Mini Protean system. The protein that is being analyzed today is the MBP-intein fusion protein that has already been purified and analyzed.

Procedure

Using the protocol from [1], the stain for the gel electrophoresis was created using 0.25g of Coomassie Brilliant Blue R250 in a 90 mL solution of methanol:H2 (1:1 v/v) and 10 mL of glacial acetic acid. It was then filtered to remove any particulate matter. This was used and applied for the Bio Rad Mini Protean System for protein expression.

A 12% discontinuous polyacrylamide gel was created using about 10 mL of resolving gel in a BIO-RAD set up. The gel was then layered with a thin amount of methanol and set for 20 minutes, after which it was removed with chromatography paper and stacking gel solution was added. After the stacking gel was set, the comb was removed from the BIO-RAD set up and a ladder and three samples from Group 2's purified Maltose Binding Intein protein were added to the wells. The gel was run for about 40 minutes and left in the staining solution overnight. It was then placed in a destain solution

Results

Gel Results.jpg

This picture shows that the protein from this experiment was not pure.

Notes

There was some gentle tearing of the gel during its removal from the BIO-RAD set up.