How to purify sample before running for LC and what internal standards can be used in LC/MS analysis of biological samples such as serum free media?
I am looking for methods which can be used to purify serum free media without losing the protein.
Hi,
You could consider using diafiltration units with a 3 kDa, or 10 kDa, molecular weight cutoff, assuming that the components you want to get rid of are small (smaller than your average protein).
With regard to internal standardization: you could spike a known amount of protein that is not expected to be in your sample, but represents the protein(s) you're interested in (i.e. casein, ovalbumin, etc.).
Hope this helps!
Thank you Eef. In addition, after obtaining my purified samples, do I need to have an additional purification step too? Before i run my analyte with the mobile phase?
If you're planning to run reversed phase, you can get rid of salts and other interfering constituents during the first couple of minutes of your gradient, so I guess if you clean the sample using the diafiltration method described above, you're good to go!
Are you thinking of analyzing your protein(s) intact, or after proteolytic digestion?
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