I did protein purification from polyacrylamide gel because i want to cut only target size. But, after running SDS for checking purification, there are several bands were shown. Moreover, there are bigger size than my target band. If you could know please give me some suggestion to solve this problem.
You may be getting higher order aggregates: dimers, tetramers. These may have formed during the purification process. You may need a dissociation step before running the purification check gel.
You may also want to check your protein sequence for cysteins. If there are multiple, disulfide bond formation could be a problem. Adding reducing agents like 2-mercaptoethanol or DTT can help prevent this. Glycerol can also be added to help protein stability. Regardless of the purification method, I find a good starting resource is the QIAexpressionist manual by Qiagen.
Katrina Good Thank you for your recommendation, I will try further.
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