Mouse and human caveolin-1 are very similar but can you transfect inter-species and hope to analyse effects? Western blot and immunofluorescence antibodies cross species readily, but which Taqman primer for PCR do you use - human or mouse caveolin to detect expression?
This is the clone to be used: http://www.sinobiological.com/CAV1-cDNA-Clone-g-17206.html
Yes you can.. For the primers you have to use the human one.. Even if to perform a pcr after a transient transfient is enough unusefull.. Different if you want to generate stable cell line. In this case the pcr is essential... However,I transfect human protein in mouse cells and vice versa evrery day..no problem.. Good luck
Degrada,
Francesco
Thanks Francesco - do you gave a reference (published article) with such a human DNA transfection in mouse cells please. Also, can you overexpress a human protein with this transfection, on top of the mouse protein?
Dear Sourabh,
I was co author in a paper in which I trasfected the human gene in MEFs cells. I don't remember if it is wrote in material and methods but you can be sure that there aren't problems for this kind of transfection. I don't understand very well your new question... If you want to be sure that you are following the effect of the transfected protein you have to kd the endogenous one. You can avoid to silence the endogenous gene only if you want to see if your esogenous protein can act as dominant negative. In over espression condition, for sure, you have a lot esogenous protein but the endogenous is always present and you can't say nothing only because there is a lot of transfected one.
Regards
Francesco
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