Western Blot: Technique, Theory and Trouble Shooting

Dear Editor,

We appreciate the comments[1] regarding our published article “Western blot: Technique, theory, and trouble-shooting”.[2] For transferring of proteins to a membrane, the method of “wet transfer” in Western blotting is routinely utilized by laboratories and guided by the protocols online.[3] Heat may be produced during the transfer; according to the recommended protocols, placing the transfer apparatus on ice or placing it in the cold room is necessary. It should be considered that the purpose of ‘Figure 7’ in our article[2] is to show how to cool down the environmental temperature to 4°C by placing the apparatus on ice, which cannot be misunderstood as submerging the electroporator or the power leads in the water bath. In addition, we agree with the proposal that safe laboratory practice should always be observed to avoid chemical or electrical hazards in operating the Western blot transferring systems.