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Far western blotting

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5 replies to this topic

#1 Divya Pothula

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Posted 01 April 2012 - 06:35 PM

Hello everyone!

I am trying to work on some protein-protein interactions, and have recently read a few articles and papers that Far western blotting is a very good technique to identify them.
Has anyone used this technique before and could help me describe it for me so that I can get a fair idea on how to do it please?

Thanks
D

#2 Inmost sun

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Posted 02 April 2012 - 10:13 AM

it is similar to immnoblot but w/o non-antibody protein; recombinant protein to be tested may carry a tag to be detected by antibody, or you use an antibody directed against bound protein

#3 mdfenko

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Posted 02 April 2012 - 10:33 AM

see this wikipedia explanation: far western blotting.
talent does what it can
genius does what it must
i do what i get paid to do

#4 Divya Pothula

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Posted 03 April 2012 - 04:42 PM

Thnks all! I am gathering information on it and trying my first trial soon. ;)

#5 Inmost sun

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Posted 04 April 2012 - 02:52 AM

Good luck! I started FWB with radioactively autophosphorylated (purified) kinase; it worked very well but a control for non-phosphorylated form was lacking; so I went ahead with antibodies....

#6 Divya Pothula

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Posted 09 April 2012 - 06:04 PM

Thanks I have not tried this yet...but then I am very much willing to...as I recently started my PhD...I am firstly trying to purify my recombinant protein and then tag it but not radioactively..some other options... well still working on these.




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