Why voltage cannot be set in wet western blot transfers?
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Apart from an explanation of Ohms law describing the relationship between voltage current and resistance, the most relevant details are to be found in Jurgens answer
I will to what he said by saying that when resolving different sized proteins by PAGE the use of a constant voltage or current is not so critical
The more crictical part oif the procedure is subsequent transfer of proteins to NC or Hybond, i.e. blotting. Genrally speaking you run at a constant and usually low current to minimise the generation of heat as this can complicate blotting efficiency and lead to the gel sticking to the NC (or hybond) if the current is too high and therefore too much heat is being generated. To elaborate in terms of Ohms law, the higher the voltage the higher the current for a given resistance (which admittedly changes during blotting owing to electrolytic break down of your blotting buffer): this leads to more effective and rapid transfer but generation of more heat and associated problems (described). However, even this can be controlled at higher currents for more rapid transfer by simply immersing the blotting rig in ice or performing blotting in a cold room (4C).
I will to what he said by saying that when resolving different sized proteins by PAGE the use of a constant voltage or current is not so critical
The more crictical part oif the procedure is subsequent transfer of proteins to NC or Hybond, i.e. blotting. Genrally speaking you run at a constant and usually low current to minimise the generation of heat as this can complicate blotting efficiency and lead to the gel sticking to the NC (or hybond) if the current is too high and therefore too much heat is being generated. To elaborate in terms of Ohms law, the higher the voltage the higher the current for a given resistance (which admittedly changes during blotting owing to electrolytic break down of your blotting buffer): this leads to more effective and rapid transfer but generation of more heat and associated problems (described). However, even this can be controlled at higher currents for more rapid transfer by simply immersing the blotting rig in ice or performing blotting in a cold room (4C).
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