6. How do I avoid "splotchy" or uneven Western Blots?

6. How do I avoid "splotchy" or uneven Western Blots?

To avoid "splotchy" or uneven blots when performing a Western Blot analysis:
a. Extend the blocking time to optimize blocking effectiveness.
b. Extend wash cycles (this is particularly important for tissue homogenate samples).
c. Make sure that milk powder is in solution completely before adding it to the blot.
d. Spin the tube containing the antibody solution before removing some for use, in case of protein aggregates.

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