What Is the Purpose of Keeping Tissue Homogenate on Ice During Total Protein Extraction in Western Blot?

In Western blot experiments, briefly placing the tissue homogenate on ice following homogenization serves a critical role in preserving protein integrity. This step primarily aims to prevent thermal denaturation of proteins caused by heat generated during mechanical disruption. Maintaining the homogenate at low temperatures helps stabilize the native conformation of proteins and minimizes the activity of endogenous proteases and phosphatases, thereby improving the accuracy and reproducibility of downstream protein analysis.

 

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