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Precipitation during dialysis

dialysis issue

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#1 Hong Zhan

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Posted 22 June 2012 - 10:33 AM

Hi everyone,

Recently, I did some dialysis for coupling reaction. The IgG (protein used for coupling dye) is in PBS buffer contain 30% glycerol. When I added NHS-SS-biotin in IgG solution and finished coupling reaction. I did dialysis against PBS to remove exceed biotin reagent. But I found precipitation in the dialysis tube (from spectrum lab). All process was done at 4C.

My hypothesis is glycerol crystalization??? I am not sure if it is my IgG protein..

Anyone have any idea???

I will run western blot tomorrow...

Thanks!!!

Best,

Hiroshi

#2 Hong Zhan

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Posted 22 June 2012 - 11:39 PM

I think I now .... the antibody I bought is not 100% pure. Maybe it's other protein in the solution precipitated during the dialysis

#3 Inmost sun

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Posted 07 July 2012 - 04:09 AM

unlikely glycerol precipitated; maybe the antibody: solubility was decreased because of cooling AND loss of glycerol




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