What would cause the "doublets" that I'm seeing on my 2D gels?
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Posted 09 March 2016 - 06:57 AM
Posted 09 March 2016 - 12:55 PM
i would think that would be proteins with similar pI and a little different molecular weight.
you can check them on mass spec.
Edited by mdfenko, 09 March 2016 - 12:56 PM.
talent does what it can
genius does what it must
i used to do what i got paid to do
Posted 14 March 2016 - 01:30 AM
I remember reading somewhere (but can't find it now) that it can be an artifact. Due to placement of the strip on the gel?
Posted 14 March 2016 - 01:42 AM
Ah, I just found something, although it's still unclear why it happens.
Posted 28 March 2016 - 02:29 PM
My GUESS (from very limited background of OChem) is that the sulfur in the thiourea reacts with a population of the protein, on the cysteine, to form a "post-translational modification" disulfide bond that modifies the molecular weight of the protein.
What is the concentration of thiourea in your sample? Any chance of calculating the molar ratio of thiourea/protein to see if we're using all the thiourea up and have excess protein?
Admins feel free to tell me I'm crazy.
Edited by Michael Starr, 28 March 2016 - 02:30 PM.