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protein extraction for use in ELISA

protein extraction for use in ELISA - Protocols and Methods Forum

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  #1  
Old 01-02-2008, 07:42 PM
James Munro
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Default protein extraction for use in ELISA



Hello,

I am hoping for some suggestions for a protein that I am trying to extract for detection in an ELISA.

I have a protein that is 22 kDa which is soluble in sodium borate at either a low pH (<4) or high pH (>9). In its pure form I can detects it in a ELISA when using specific antibodies. When the protein is in commercial fish feeds I can not longer detect it.
Has any one got an extraction method that I could try?

Thanks
James
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Old 03-01-2011, 01:16 PM
Pipette Filler
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Default Re: protein extraction for use in ELISA

Try separating your protein of interest with [Only registered users see links. ]. Then cut the bands from the gel and extract the protein with either electroelution using a [Only registered users see links. ] or digest the protein using an [Only registered users see links. ].

If you still have trouble with your sample after SDS PAGE followed by electroelution or in-gel digestion, try cleaning up the sample with a C8 reversed phase product. I use [Only registered users see links. ]. Hope this helps, good luck.
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