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Hong Zhan

Member Since 22 Jun 2012
Offline Last Active Jul 19 2012 06:04 AM

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Topics I've Started

Troubling for antibody biotinylation

19 July 2012 - 06:07 AM

Hi Everyone,

I recently did antibody biotinylation with pierce NHS-SS-biotin reagent. On western blot, antibody seemed to be biotinylated. i tried to find a way to quantify the biotinylation level.

Just any one know roughly how much biotin will be on one antiboy/??? (mouse IgG monoclonal antibody.)

Thanks.
Hong

Precipitation during dialysis

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