Five common factors affecting protein transfer efficiency

June 17, 2012 at 2:05 pm Leave a comment

If you are interested in obtaining the best protein transfer results, it is important to understand the relationship of the various factors involved. These factors include: voltage (V), current (I),resistance (R) and power (P).

Rapid protein transfers are generally obtained by applying maximum voltage while minimizing the distance betweeen electrodes. However, if the transfer conditions are inappropriate, poor transfer efficiency can result.

Two basic equations are important in electrophoresis.

The first is Ohm’s law: V = I x R

The second is the power equation: P = I x V = I2 x R = V2/R

While voltage and current are determined by the user and the power supply settings, the resistance is inherent in the system. It is therefore important to know which transfer conditions affect the resistance of the transfer system.

Below are the top five factors affecting resistance:

  1. Alterations to buffer composition; that is, addition of SDS or changes in ion concentration due to addition of acid or base to adjust the pH of a buffer
  2. Gel pH, ionic strength, and percentage of acrylamide, especially if the gel has not been properly equilibrated
  3. Number of gels (current increases slightly as the number of gels increases)
  4. Volume of buffer (current increases when volume increases)
  5. Transfer temperature (current increases when temperature increases)

For more great information, be sure to download the Protein Blotting Guide from Bio-Rad Laboratories.

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