Hello, i am currently doing a research which involves the use of proteins,buffers and HPLC. I am planning on using acetate buffer for the entirety of the research. However, the researches i am referring to, use different buffer systems at different times. I was wondering what the importance of using different buffer systems when using proteins?
Dear Isaac,
This depends on what the particular experiment is; if you are studying proteins to find out about their function, interactons with other proteins, or how they break up under physiological conditions, you will need to use a buffer that is mimicking protein behaviour in such conditions.
If you are interested in unfolding, denaturing the protein, you will need to use buffers that are probably supplemented with detergents and that may be acidic.
Best,
Hediye.
Hello,
Thank you for your response. To be a little more specific I am trying to study LDH-LDH interaction using HPLC. Do you think an acetate buffer would be a good system?
Is it a reverse-phase HPLC? Are you running the HPLC with basic buffers? what is the pH of the buffer?
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