What Are the Possible Causes of Protein Band Degradation in Electrophoresis?
If protein bands appear degraded during electrophoresis, several factors may contribute to this issue:
Improper Protein Sample Handling
Proteins may undergo degradation due to protease activity during extraction, storage, or handling. To prevent this, protease inhibitors should be added during extraction, and all procedures should be conducted at low temperatures to minimize proteolysis.
Suboptimal Electrophoresis Conditions
The stability of proteins can be affected by electrophoresis parameters such as voltage, current, and buffer pH. Excessive voltage may cause thermal denaturation, leading to degradation, while inappropriate buffer pH can alter protein stability.
Excessive Sample Loading
Overloading the sample can result in protein aggregation, leading to diffuse or smeared bands, which may be misinterpreted as degradation.
To address this issue, it is recommended to systematically assess and optimize these parameters. Additionally, testing different protein samples or modifying electrophoresis conditions can help pinpoint the specific cause of degradation.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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