Problems with AU1tag

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Thu Feb 8 09:07:33 EST 2001


This message has been posted by:  "Luis M. Martins" <L.Martins at REMOVE-THIS-TO-SENDicrf.icnet.uk>

Hi I have tried to insert an AU1 tag either at the N- or C-term of my
protein by PCR. I have inserted the sequence met-DTYRYI-GG-ORF or
ORF-GG-DTYRYI-end into pcDNA3. I have sequenced and things seem OK. I tried
to check for transient expression in 293 cells.

I was very surprised not to detect any protein by western using an anti-AU1
Ab. My control with a previously AU1-tagged protein worked (this construct
wasn't made by me)

Any thoughts?

Thank you

Miguel

-- 
Luis Miguel Martins, Ph.D.
Signal Transduction Laboratory
Imperial Cancer Research Fund
44 Lincoln's Inn Fields
London WC2A 3PX
UK 






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