As a scientist, do you ever perform biochemical experiments with inadequate buffers which limited the impact of your research? Do you face buffers with cell toxicity and could not support enzymatic activity? Do you hope to make a discovery that will change how you think about a biological process? Do you wonder which Biological Buffer is your suitable one? If yes, please check tips below to see if I can help you a bit.
Tips to follow:
- The pKa Value: The pKa value of a buffer should be less influenced by the buffer concentration. So, a normal pH value should be between 6-8 for most biochemical experiments.
- Water Solubility: The best quality buffers should be soluble in water freely. With higher water solubility, it is easier to prepare concentrated stock solutions.
- Permeability: Normally buffers should be exclusive by biological membranes . For example zwitterionic buffers include MOPS and HEPES, they do not pass through bilogical membranes. But Tris buffer has a relatively high degree of fat solubility and may therefore permeate membranes. So Tris does not isomerize into zwitterions.
- Minimal salt effects: The buffer components should not interact or affect ions involved in the biochemical reactions being explored.
- Ionic complexes: Good buffers should have very low metal-binding constants and are particularly suited to investigate metal-dependent enzymes (Good & Izawa 1972, Blanchard 1984)
- Degradation: Good Buffers should be inert or in other words “stable”. It should not react with metabolites or other components. It should not resemble with enzyme substrates.
- Absorption: Good buffers should not absorb any visible light or we can say the UV absorption should be less than 230nm. For example, the Tris buffer UV absorbance at 280nm is less than 0.1% and our tested result is around 0.04%.
- Easy and Less expensive: When you are going to do an experiment, cost for the materials will affect your planning. So choose the correct material and make your preparation and purification easy and less expensive.
Finally, wish you a good experience in souring material and smoothly experiment.
References:
Arduengo, P.M. (2010) Sloppy technicians and the progress of
science. Promega Connections
http://promega.wordpress.com/2010/03/15/sloppy-technicians
Ferreira, C. M., Pinto, I. S., Soares, E. V., & Soares, H. M. (2015). (Un)suitability of the use of pH buffers in biological, biochemical and environmental studies and their interaction with metal ions – a review. RSC Advances, 5(39), 30989-31003. doi:10.1039/c4ra15453c.
