Overview: [Characterization of red cell membrane proteins by SDS-PAGE] [research paper]
Topics: [gel analysis] [molecular mass standard curve] [measuring relative mobility] ["Hall of Shame"]
Data: [gel images]

Bizarre Results Explained

Bizarre – example 1

The large container of glycine looked the same as the large container of choline chloride. Choline is a lot more hydroscopic than glycine, which should have been a clue that the wrong component was used in the electrode buffer. Choline is a basic compound and certainly does not substitute for glycine. We caught the error and replaced the buffer after electrophoresis had proceeded for some time, but it was too late for this one.

Bizarre – example 2

This one was kindly contributed by T. Sedlacek, Czech Republic. It was run with high current and overheated.

Bizarre – example 3

Here are two gels that were contributed by E. Morales Rayas. A likely explanation is that each time there was a delay between loading the samples and actually running the gel. The middle lanes of the first gel show an alternating pattern in which every other lane is either wide or compressed. Bands on the second gel spread out toward the edges.

When samples sit in a well the proteins begin to diffuse into the stacking gel, both vertically and laterally. Smaller proteins diffuse more rapidly than do larger ones. If proteins in a sample diffuse laterally they may alter the electric field affecting adjacent lanes, especially if the samples in adjacent wells contain predominantly higher mass polypeptides. The problem isn't so bad when all of the samples are of similar composition (second gel) and the lanes are loaded in sequence. However distortion still occurs.

Bizarre – example 4

Evidently it is not a good idea to store tris-glycine electrode buffer in a gallon plastic bottle on a shelf in the lab. We didn't see any mold in the buffer, but by golly it was there, all right. Notice that in each example the gel below the dye front was cleared following destaining. The persistent blue background is from mold proteins that continued to penetrate the gel throughout the procedure.

Bizarre – example 5

Here's one thing that can happen if someone opens a gel box before the run is completed, WITH THE POWER ON! Another thing that can happen is that the bozo can get killed! This one was contributed by Margaret A. Buttery, Notre Dame De Namur University in Belmont, California.

 

 


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Created by David R. Caprette (caprette@rice.edu), Rice University 26 May 05
Updated 28 Nov 06