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protein biotinylation


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#1 claire84

claire84

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Posted 16 September 2014 - 04:39 AM

Hi everybody,

I have some problem with the biotinylation protocol for a protein and I need your help.

I bought from Pierce a "Sulfo-NHS-LC-biotinylation kit" and I used it to incorporate the biotin into my protein (fibronectin).The kit I bought was for the biotinylation of 1-10mg of protein as weel as the protocol. 

However, I used it to target a protein ammount inferior to that reported into the protocol. Maybe for this reason I encountered some problem regarding the incorporation. After the quantification of mol of biotin per mol of protein, I determined a great ammount of biotin into the sample. 

 

Something goes wrong in the purification step?How can I check the quantity of unreacted biotin in my final sample?

 

Do you have some experience with this kind of biotinylation kit?Please find attached the file of the biotinylation protocol.

 

thank you,

 

Chiara






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