I used Y-PER protein extraction reagent to extract yeast protein. Then, I quantified the amount of protein by BCA. The problem is when I using this protein to run SDS-PAGE, the loading dye in gel looking very weird. It dispersed around in the gel after running ( not a sharp band/ look like a band at all.
I then checking by staining with Cromassie. It turned out that there is no protein at all. I do not know how to figure this. Anyone who has an experience on this please help or any suggestions.
Picture is useful - I m not sure what you are talking about the gel. Concentrate / try to load more protein
Mahesh Chandak Thak you for the answer. I will concentrate the protein and try again. Have you used Y-PER with SDS-PAGE before?Mahesh Chandak
What do you mean by it dispersed around the gel? It was during running or after running? Had your protein ladder migrated as it should? What were the results or your BCA assay? How much protein did you load per well?
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