Hello all!
I am a student at CPCC and am doing a project dealing with Adipocyte Protein 2. I was able to discover the pi value and the isoelectric point. Thus giving me the pH value of the buffer needed (6.5). I was wondering what type of buffer solution would you recommend for this. (I will be doing a Diafiltration method for this project.) I feel at a loss for what chemical or chemicals I should use.
I was looking at this website (https://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/lysis_buffer_additives/) for some help, although much information could be found, I still could not decipher what I should use as I have very little experience in this situation.
I hope I am looking at this all correct. Thanks for any assistance you my offer,
Micah L
https://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/lysis_buffer_additives/
A useful approach is to read papers published on this protein that investigated the fatty acid binding characteristics of the purified protein. That should give examples of buffers that others have found useful.
By the way, the isolelectric point (pI) is not a useful guide to the optimal buffer pH for functional measurements of a protein. People often try to avoid putting a protein at its pI because there is a risk of precipitation due to the lack of a net charge.
Thanks for your response. I will look into this further. I appreciate you bringing up the pI and the risk of precipitation. As you can tell this topic is very new to me.
Thanks again,
Micah
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