What Are the Possible Causes and Solutions When Proteins Fail to Migrate During Western Blot Electrophoresis?
When a Western blot yields no observable bands or fails to produce detectable results, several technical issues may underlie the problem:
Power Supply Configuration
1. Verify that the electrophoresis power supply is properly connected and operational.
2. Ensure that voltage and current settings are appropriate for the gel type and expected run time.
Electrode Polarity Orientation
Confirm that the electrode connections are correctly polarized. Reversed polarity may prevent proteins from migrating toward the correct electrode, resulting in no band formation.
Sample Quality and Integrity
1. Assess the purity and concentration of the protein samples. Inadequate purity or excessively dilute samples can lead to undetectable or faint signal bands.
2. Evaluate storage conditions. Samples that have been repeatedly freeze-thawed or exposed to ambient temperatures may undergo protein degradation or functional loss, impairing detection efficiency.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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