Antibody Biotinylation

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Antibody Biotinylation

Postby jonathan07 » Apr 30 2016 2:27 am

Hello,
I'm not use to do immunochemistry but I would like to biotinylate 500 microgrammes of an 2.4mg/mL antibody which means a volume of 208 microlitres.
To proceed to the biotinylation I will use the thermofisher kit n.21327 ( https://tools.thermofisher.com/content/ ... tin_UG.pdf) but there is something in the procedure that I don't understand concerning the part for proteins in solution:

I need to prepare the antibody in PBS according to calculation in Section A. I came on an volume of Biotin to add of 6.65 (6.7) uL for 208uL of Ab's but how much PBS do I need to add ?
Maybe it's a very basic question for some of you but if someone could please give me a hand , I would very appreciate it.

Thanks,

Jonathan
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Re: Antibody Biotinylation

Postby mdfenko » May 02 2016 3:27 pm

in what buffer is your protein? if it is amine-free and within pH 7.2-8.0 then you don't have to exchange with pbs.

the calculation given in part A is to determine the moles of protein and how much biotin to prepare a 20-fold excess of biotin to protein (mole/mole). final reaction volume is not specified and, therefore, is probably not important (other than to ensure that you add enough biotin).
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Re: Antibody Biotinylation

Postby JaneSmithSkymeter » Jun 29 2016 9:09 am

Some companies provide antibody biotylation and the quality is high, you can search it on website.
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