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-   -   Ion exchange chromatography - Cation exchangers and TRIS Buffer? Compatible? (http://www.molecularstation.com/forum/biochemistry-forum/84887-ion-exchange-chromatography-cation-exchangers-tris-buffer-compatible.html)

Valine 03-11-2012 12:35 PM

Ion exchange chromatography - Cation exchangers and TRIS Buffer? Compatible?
 
Hi People,

thats more or less my problem,

I'm having some problems to purify my peptide by reversed phase chromatography. I have decided to move to Ion exchange chromatography since my peptide is quite basic (it has Lysine, Arginine). Actually the sequence is like KARKSAGA where in the K in the middle of the sequence it has been modfied. The pI of my peptide is around 10-11. So I have decided to use cation exchanger. The problem is that my peptide is dissolved with Tris buffer and I would like to use also as eluent Tris buffer. So far I read that Tris buffer is not recommended for Cation exchangers...

My question is Why?

Thanks

qianhaha 06-06-2013 04:44 PM

Re: Ion exchange chromatography - Cation exchangers and TRIS Buffer? Compatible?
 
In science, peptide synthesis is characterized as the formation of a peptide bond between two amino acids. While the definition of a peptide is not definitive, it usually refers to flexible (little secondary structure) chains of up to 30-50 amino acids. <a href="http://www.unitedpeptide.com">United Peptide</a> provide peptide synthesis services.

The ability to form peptide bonds to link amino acids together is over 100 years old, although the first peptides to be synthesized, including oxytocin and insulin, did not occur for another 50-60 years, demonstrating the difficult task of chemically synthesizing chains of amino acids (1). In the last 50 years, advances in protein synthesis chemistry and methods have developed to the point where peptide synthesis today is a common approach in even high-throughput biological research and product and drug development (2).

The benefit of peptide synthesis strategies today is that besides having the ability to make peptides that are found in biological specimens, creativity and imagination can be tapped to generate unique peptides to optimize a desired biological response or other result. This page highlights the important aspects of peptide synthesis, the most common methods of synthesis and purification and the strengths and limitations of the respective strategies.


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