My TA for an SDS-PAGE lab stained our gels for us and ended up with the following result. He used Coomassie blue for 30 min and then he left it overnight in an unknown destaining solution. Does anyone know where he went wrong? We ended up with a blue gel and no definitive stain or bands
Did you stain using the rapid coomassie blue that can be detained with distilled water?
Looking at them I think the stain did not cover the gel and so parts dried. If you can provide more info on the stain type and destaining solution constituents it will make troubleshooting better.
Franklin Yengdem Nuokpem So sorry I just updated my answer but yes he did use coomassie blue and then the lab report just says destaining solution so I am unaware of what that solution was actually made of but to my understanding, it required being done in a fume hood.
For one thing, from the looks of the photos, it seems that there was just not enough destaining solution and the gels were left to dry out.
Ok, I think he used the rapid coomassie stain which can stain in less than an hour. For that, the destaining solution is usually distilled water and it is not usually for very long; not overnight. I think he used destaining solutions with a high percentage of acetic acid which is usually for in house prepared coomassie stains that are used overnight or more than 1 hour. Aside from the gel looking dried which is reversible by adding distilled water and incubating at room temperature for about an hour; I think you should restain the gels again using the staining and destaining protocols on the rapid coomassie bottle. I think the overnight destaining cleared the stains from the bands.
Alternatively, you may do a ponceau S staining which gives results in less than an hour to be sure whether you have protein bands in there or not. Cheers.
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