What is the recommended method for routine cleaning of a cation exchange column?
The recommended method is to rinse the column with 100% water for 30 times the column volume for desalination, and then store it in a buffer.
What should be used to wash the strong retention of contaminants in a cation exchange column?
A high-concentration buffer salt solution (5-10 times that of the mobile phase, but not more than 80M) should be used to wash the strong retention of contaminants.
What should the sample solution for cation exchange chromatography not contain?
The sample solution should not contain any particulate matter.
What should be done with the mobile phase's pH value in cation exchange chromatography?
The pH value of the mobile phase should be controlled at 2-8.
Can the mobile phase be prepared in advance for cation exchange chromatography?
No, the mobile phases are best prepared on the day of the experiment.
What should be avoided with the ion column?
The ion column should not be dropped or touched.
How does a protein's net surface charge change with pH in cation exchange chromatography?
A protein's net surface charge changes with pH based on its isoelectric point (pI). At a pH equal to the pI, the protein carries no net charge. Below the pI, it carries a net positive charge, and above the pI, it carries a net negative charge.
How is a buffer chosen in cation exchange chromatography to ensure a known net charge for a protein of interest?
A buffer can be chosen based on the protein's pI, ensuring that it carries a net charge of interest.
How are proteins separated in cation exchange chromatography?
Proteins are separated based on their varying degrees of charge at a given pH. Proteins with different pI values will have different affinities for the charged surface groups on the resin particles, allowing for their separation.
What is used to elute proteins in cation exchange chromatography?
A salt gradient is used to elute proteins, with proteins eluting in an order depending on their net surface charge.
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