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Different results invitro and invivo


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#1 jaya2020

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Posted 09 January 2015 - 07:08 PM

Hi,

Iam working with the expression of Fgf 23 in osteoblast and cDMP1. invivo published results prove that cDMP1 inhibits Fgf23. But when I studied the promoter activity of fgf23 invitro using cDMP1 by duel luciferase assay, I see that it increases the promoter activity of Fgf 23 rather than inhibiting it, which is directly contrast to what is reported in vivo. what could be the reasons for this. Im confused.

 



#2 bob1

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Posted 11 January 2015 - 12:12 PM

Perhaps the interaction is not at the DNA/RNA level but at the protein:protein level.



#3 jaya2020

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Posted 12 January 2015 - 07:42 PM

Thanks Bob. I would like to know how can I study it. Iam in need of some help and this is the first time for me to do this work



#4 bob1

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Posted 12 January 2015 - 11:55 PM

The usual way is to determine if the proteins interact is by doing co-immunoprecipitation - i.e. precipitate using an antibody against cDMP1 and then try detecting with a Fgf antibody. Of course you need appropriate controls (non-specific antibody or normal serum of the same species as the precipitating antibody) to determine if the interaction is real. You can also do in-vitro expression of the proteins and see if they form a complex



#5 jaya2020

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Posted 13 January 2015 - 12:02 PM

Thank you, I will try it out.






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