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Receptor discovery/cloning


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#1 cupidstunt

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Posted 21 April 2009 - 04:22 AM

Hi all,

I have a basic question. I was wondering how do they discover receptors? If you know there might be a receptor for a known ligand, how do u sort of purify it? sequence it? amino acids sequence etc?? any ideas??

thanks a lot!!

#2 perneseblue

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Posted 21 April 2009 - 04:56 AM

This isn't my field. But the traditional thing to do is to bind your ligand to a column and run a protein extract through the column. Proteins that do bind to the ligand will be retained. You can dissociate your protein from the column and with some work you can then isolate a pure sample of your protein or proteins (perhaps using HPLC or 2D gel). The protein AA sequence can then determined using mass spec or an Edman degradation sequencer
May your PCR products be long, your protocols short and your boss on holiday




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