After tamoxifen induction (5 DAYS IP) cre recombinase is not trans locating to the nucleus.

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Yuan-Yeu Yau

How do you know the Cre protein did not translocate into nucleus?

Manu Prasad

I did immunofluorescence of mice tissues with Cre antibody after 5 days of tamoxifen treatment.

Did you include a positive control tissue in your immunofluorescence experiment?

By the way, I checked online:

Is your KRT14-cre mouse the same as this one: Tg(KRT14-cre)1Amc/J (from The Jackson Lab, https://www.jax.org/strain/004782)? If so, the promoter is a human promoter (human keratin 14 promoter). Had you replaced the promoter with an inducible promoter?

Thank You

I don't have any positive control. STOCK Tg(KRT14-cre/ERT)20Efu/J.

https://www.jax.org/strain/005107.

If you don't have a positive control, how do you know that the negative results from immunofluorescence assay is not due to assay procedure problem?

I have mouse embryonic fibroblasts with stable cre expression.It was in the nucleus.

What will you suggest as a positive control.After tamoxifen induction cre-ert should move to the nucleus.So I was expecting cre in the nucleus.

I don't know it is the problem with tamoxifen administration.

Thanks