Good day, I have seen that our laboratory has a lot of stock of Precision plus unstained standards and very few of Precision plus stained standards.
Since I need to run a lot of western blots in the future, I was wondering if I could stain the unstained ladder BEFORE running gel electrophoresis and load it on the well already stained.
I read online that you can stain the unstained ladder with coomassie blue. I was wondering, can you stain it with commassie blue, load it on the wells and I can have my ladder?
The catalogue number for the unstained ladder is: 161-0363
Eduardo,
I personally have never tried this before but it would be a quick experiment to test out, otherwise you can run a non-stained marker and check the gel with commassie blue or S-Ponceau stain
Hello Eduardo,
For analyzing the western blot results you need a stained ladder for determining the precise molecular weight of gained band(s).
At first I recommend you to use pre-stained ladders because these ladders have no expensive costs.
However, I didn't test the protocol you describe before but I think it doesn't work well. I think the commassie blue-protein complex will be destructed during SDS-PAGE gel running or during protein bands transfer onto the blot paper.
If you want to do with unstained ladder yet, I recommend to do all steps of western blot just routinely do. After transferring the protein bands onto the blot paper, stain the paper with Ponceau-S for visualization of ladder. Then, cut a small piece of the paper standing in front of your desired protein band. After this, you can complete the western blot steps and analyze your desired band at the end.
Note that do not use pencil marker for determining your desired band.,because it will be lost during washing steps.
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