[Protein-analysis] Setting up an ELISA: any advice gratefully received!

Stearns, Adam T via proteins%40net.bio.net (by ASTEARNS from PARTNERS.ORG)
Mon Feb 19 15:09:20 EST 2007


Help!

I wonder if anyone might be able to give me some advice about how to go about
setting up an ELISA: I'm looking at fairly subtle changes in membrane protein
expression (SGLT1 transporter) in intestinal tissue samples. I have been doing
Western blots, and quantifying with densitometry, but I want to try to get
slightly more quantitative results. My initial main concern is the protein
extraction. Up till now, I've been using triton X in lysis buffer. Will this
interfere with the ELISA, and do I need to find a different extraction buffer
for extracting the protein? Alternatively, does anyone know if I can remove
detergent from the samples?

With many thanks!

Adam Stearns
Boston, MA
US.





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