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Can TCA percipitate Glycoprotein?


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4 replies to this topic

#1 arctos

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Posted 05 April 2010 - 11:44 PM

Dera folks
Can TCA and aceton percipitate glycoprotein? The proteins including glycoproteins are in 15% sugar solution. I want to isolate the total protein.
And did anyone use the total glycoprotein isolation kit from Qiagen. Is it efficient or not. I am worried about the sugar in the solution might interfere the glycoprotein binding onto the immobilized lectin. Any suggestions?

#2 K.B.

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Posted 06 April 2010 - 02:16 AM

It should work, although I don't know how would sugar behave. Add TCA to sugar solution (without protein) to see what happens.

I don't know what principle this kit is based on - if it's lectin based, sugar in solution may interfere. Kit manual should give you acceptable limits of additives in sample. Anyway, it's not a problem - you may simply dialyse your sample to remove sugar.

#3 mdfenko

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Posted 06 April 2010 - 06:39 AM

sugar makes the solution dense so it will be difficult to pellet the precipitate. as recommended by k.b., you should dialyze the solution.
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#4 arctos

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Posted 06 April 2010 - 05:05 PM

Thank you for your replies. I tried dialyze the solution. As the protein conc. is only 20ug/ml and mostly are glycoprotein, they could bind onto semi-permeable membrane. I lost a lot. Do you think whether pretreating the membrane with tween 80 can save a little or not?

#5 mdfenko

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Posted 07 April 2010 - 07:14 AM

adding salt (0.15-0.2M) to the solution should prevent binding to the membrane.
talent does what it can
genius does what it must
i do what i get paid to do




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