Why Do Western Blot Bands Appear Inconsistent for Proteins Extracted from the Same Batch?
When conducting a Western blot experiment, inconsistent band patterns from the same batch of extracted proteins may arise due to the following factors:
Variations in Electrophoresis Conditions
Fluctuations in voltage or current can impact protein migration and separation.
Gel-Related Issues
Defects in the SDS-PAGE gel, such as uneven polymerization or prolonged storage, may affect protein resolution.
Transfer Process Irregularities
Non-uniform current distribution or improper transfer time and voltage settings can lead to inefficient or uneven protein transfer.
Antibody-Related Factors
Variability between antibody batches or antibody degradation due to expiration can result in inconsistent detection.
Procedural Errors
Minor inconsistencies in sample loading, buffer preparation, or handling during the experiment may contribute to variability in band appearance.
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