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Protein stability at pH8.3


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#1 Wyanstown

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Posted 15 October 2012 - 04:24 AM

Hi,

I want to label my protein (platelet integrin 2b3a) with an amine reactive dye. The protein comes in Tris buffer (pH 7.4) which I need to replace with 0.1M sodium bicarbonate buffer at pH8.3. To do this I was going to dialyse the protein against the new buffer. Will the protein be ok for 24 hours or more in a pH8.3 buffer with 0.1 sodium bicarbonate?

Thanks

Edited by Wyanstown, 15 October 2012 - 04:38 AM.


#2 HOYAJM

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Posted 15 October 2012 - 11:08 AM

I think it will be OK. You may need to dialyze at 4C as well. You can not really predict if your protein will be "OK" based on this information. You can try to find literature about stability of this protein at different pH, or whether or not sodium bicarbonate has an effect, but the easiest way is just try it and see what happens. 24 hours is common for dialysis to change protein buffers.

Go to this site, paste your protein sequence, and determine what the charge of your protein is at these two pH (7.4, 8.3). Here you can get some insight

http://www.scripps.e...m/protcalc.html

Edited by HOYAJM, 15 October 2012 - 11:08 AM.


#3 Wyanstown

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Posted 16 October 2012 - 05:03 AM

Thanks! I'll try that




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