I need a good protocol to use for monitoring the endogenous wnt/ beta-catenin signal and activity in some cell lines I am using. Where can I find wnt agonists commercially?
Hi Jude,
monitoring Wnt/beta-catenin activity with Topflash reporters is pretty simple, as long as tranfection is working.
There are several options:
Do you need the wnt agonist just as a positive control? If so, you might consider using LiCl at final concentration between 10-40mM, or 6-BIO at concentration between 500nM and 5µM (GSK3beta inhibitor IX) to inhibit GSK3beta.
Alternatively you can buy recombinant Wnt3a, which is probably the best, but expensive, option if you want a "real" activation of the Wnt-signalling cascade. There are several companies selling Wnt3a.
Another trick is to transfect your cell with a Wnt3a or Wnt1 expression plasmid to get autocrine activation of your reporter.
If you need more detailed information let me know.
Best
Kai
Hi Jude,
monitoring Wnt/beta-catenin activity with Topflash reporters is pretty simple, as long as tranfection is working.
There are several options:
Do you need the wnt agonist just as a positive control? If so, you might consider using LiCl at final concentration between 10-40mM, or 6-BIO at concentration between 500nM and 5µM (GSK3beta inhibitor IX) to inhibit GSK3beta.
Alternatively you can buy recombinant Wnt3a, which is probably the best, but expensive, option if you want a "real" activation of the Wnt-signalling cascade. There are several companies selling Wnt3a.
Another trick is to transfect your cell with a Wnt3a or Wnt1 expression plasmid to get autocrine activation of your reporter.
If you need more detailed information let me know.
Best
Kai
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