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Western Blotting Membrane Blocking Products

membrane blocking


Nitrocellulose membranes are ideal for blotting due to their ability to nonspecifically bind proteins. This allows your target proteins to bind the membrane and become immobilized, but it also would cause your antibodies to bind the entire membrane, producing no usable data. To prevent that from happening, the membrane must be saturated with a nonreactive protein to block the nonspecific binding of your primary and secondary antibodies.


After completing the transfer of the protein sample to the membrane, thoroughly wash the membrane with Boster PBS wash buffer, then immerse the membrane in Boster TBS blocking buffer for 1-2 hours. Wash the membrane again to remove excess blocking buffer, and your membrane is ready for antibody incubation.

For a detailed protocol for membrane blocking see our Western blot handbook.


  1. Insufficient blocking can cause high background signal.
  2. Using nonfat milk blocking buffer can interfere with avadin-biotin detection systems

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