Four Great Tips for Effective Protein Blotting

January 17, 2012 at 1:02 pm Leave a comment

Are you doing western blotting or other protein blotting techniques? Checkout the following blotting tips from Bio-Rad Laboratories.

  1. Stirring during transfer helps maintain uniform conductivity and temperature. Failure to properly control buffer temperature may result in poor transfer and poses a potential safety hazard.
  2.  

  3. Electrophoretic transfer entails large power loads and consequently, heat generation. The tanks are effective thermal insulators and limit the efficient dissipation of heat; thus, simply placing the tank in the cold room is not enough to remove all of the heat generated during transfer. Effective cooling required for high-intensity field transfers and recommended for long, unsupervised runs can be provided using the cooling coil or Bio-Ice™ units included with your transfer device.
  4.  

  5. Evaluate transfer efficiency at various field strengths (V/cm), staying within the recommendations for each instrument. For transfers using high power, monitor the transfer carefully and use cooling as needed.
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  7. Perform a test run to determine the time required for complete transfer. Times may vary from 15 min to overnight, depending on many factors, including the power setting, gel percentage, and the size, shape, and charge of the protein.

 

Did you find these tips useful? For more great tips, check out the Protein Blotting Guide from Bio-Rad Laboratories.

Entry filed under: Gel Imaging and Western Blots, Miscellaneous Videos and Technotes, protein blotting, Western Blot and ELISA. Tags: , , .

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