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methods to detect EGFP expression ?! - we have a big problem! (Mar/17/2009 )

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little mouse on Mar 18 2009, 02:29 AM said:

I think I get it.
Reduce the formaldehyde concentration to 0.5-1% because it reduces GFP signal and increases background.

thanks, we'll do that next time. we're gonna do many experiments this week and hopefully we'll get the result we want.

swanny on Mar 18 2009, 03:03 PM said:

Naive question, but is the construct in the correct frame? I only ask because we had two occasions last year where non-expression of proteins came down to a (missed) frameshift.

pEGFP.N2 is a commercial plasmid from Clontech I think, so it must be in the right frame. I used the same plasmid 5 years ago and had no problem.

scolix on Mar 21 2009, 10:51 PM said:

we use lysate from a single well in 24 wells to detect GFP. Better to wait atleast 48 hrs before checking expression.

thank you.

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