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Western Blot Optimization

Written by Super User. Posted in Protein

Information on how to optimize western blots.

This is your portal for optimizing western blots!

Western Blot Optimization Table of Contents

What is Optimization of Western Blots?

Optimization of western blots is important as this is done to increase the signal to noise ratio of your results, and also decrease non-specific bands. Optimization is carried out usually in a trial and error or a more organized approach by using different blocking, incubation and washing conditions.

Factors in Western Blot Optimization

Several factors are key in optimizing Western Blots:

  • SDS-PAGE Gel electrophoresis - without a good gel you will not get a great membrane
  • Selecting the Right Antibodies (both primary and secondary) and Using them Properly (dilution is important)
  • Blocking Conditions
  • Washing Conditions
  • Development Condition

Factors in Western Blot Optimization

Several factors are key in optimizing Western Blots:

  • SDS-PAGE Gel electrophoresis - without a good gel you will not get a great membrane
  • Selecting the Right Antibodies (both primary and secondary) and Using them Properly (dilution is important)
  • Blocking Conditions
  • Washing Conditions
  • Development Condition