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Urea and protein storage


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6 replies to this topic

#1 bachai

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Posted 15 March 2009 - 07:51 PM

Can someone please tell me whether storage of a protein (inclusion bodies of a recombinant protein containing Cys, protein concentration >10mg/mL) in 8M urea at minus 80oC would result in some chemical changes? Thanks

#2 sitting_at _the _AKTA

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Posted 15 March 2009 - 08:05 PM

i store my purified protein in 8M urea at 4 degrees for ages and it seems fine, if that helps ;)

#3 T C

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Posted 15 March 2009 - 08:06 PM

Hey

I don't know about long term storage but for a short duration, just leave it at room temperature....Urea will precipitate even at 4 degrees so -80 is surely not recommended according to me.....However I never stop at this stage.

Best
TC

#4 bachai

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Posted 15 March 2009 - 08:13 PM

I'm so impressed with your super fast answers.
This helps to reassure me somehow. Great thanks.

#5 mdfenko

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Posted 16 March 2009 - 08:09 AM

a problem could arise when you defrost the solution. urea does not like temperatures above rt.

if you defrost at higher temperature (to speed it up) then you run the risk of carbamylating your protein.
talent does what it can
genius does what it must
i do what i get paid to do

#6 bachai

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Posted 16 March 2009 - 07:23 PM

a problem could arise when you defrost the solution. urea does not like temperatures above rt.

if you defrost at higher temperature (to speed it up) then you run the risk of carbamylating your protein.


Dear mdfenko. You are just terrific!
When at rush I'd thaw samples by holding/rubbing in palm instead of using an waterbath. How much carbamylating could this generate? Anyway, I will wait on ice from now.
Do you know if carbamyls are known for affecting the antigen-antibody binding? Thanks a lot!

#7 mdfenko

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Posted 17 March 2009 - 10:52 AM

Do you know if carbamyls are known for affecting the antigen-antibody binding? Thanks a lot!

it depends on the specificity of the antibody.
talent does what it can
genius does what it must
i do what i get paid to do




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