I have a protein labeled with a fluorescent dye (fluorescein) and I am hoping to get a quick measure of purity using TLC after dialysis. TLC is quick and easy so I am hoping to start there.
What types of plates and solvents should I purchase? Normal phase or reverse phase? I know I can use ninhydrin to develop the proteins but that's about it. I have had trouble finding information elsewhere...
I have never heard of separating whole proteins on TLC. I would guess it could be done using an ion-exchange stationary phase and an aqueous buffer. It turns out someone has reported this:
Article Thin-layer ion-exchange chromatography of proteins
As far as the purity of your labeling protein is concerned, the key thing is to remove the unincorporated dye. The fastest and easiest way to do that is by using gel filtration desalting, such as with a prepacked PD-10 column.
"for protein purity analysis via thin layer chromatography" ??
None. *We do not use TLC for protein PURITY analysis. It is possible to resolve apart some proteins on plates (using specialized plates), but this is a very crude measurement and not suitable for purity determination. Try HPLC.
Please research this topic before you design any experiments or projects.
Ion-exchange chromatography or HPLC will help you rather than TLC..
PAGE can also help you to check the purity of protein.
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