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SaOS2 Cell Line and P57


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#1 manuni

manuni

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Posted 02 April 2009 - 09:58 AM

Hi Friends,

I am working with p57 in SaOS2 and U2OS cell lines. Since last year i tried several time to get protein expression (western Blot) of p57 in SaoS2 cell line, but unfortunately i did'nt manage to get. Normally i use High Salt Lysate buffer with Protease Inhibitors to Harvest the cell.
I got protein expression of p57 in U2OS but not with SaOS2.

If i couldn’t manage to get this then i will be in big trouble. :D I need help so if any one knows about any literature that shows SaOS2 cells lack p57 or if any one knows how to get expression plz plz let me know

One more thing which i want to discuss is, i have tried several time to clone p57 insert (515 bp length) in PGL3 vector after cutting both with XhoI and SacI enzymes but result comes like this i got colonies but after getting DNA from Mini Preb when i cut the DNA with same enzymes they never gave me required insert. I have tried ever thing like dephosphorylation but..........

I really need some help and any suggestion will be appreciated.

Regards

manuni




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