I am new to invitro analyses. I do not know, what primary and secondary antibodies to use (which brand) for western blotting from cell line derived proteins. Can you explain in brief please?
We routinely use monoclonal antibody (clone DO-1), which is availible from different companies und works very well for endogenous p53 (tested in HepG2, Hek293, Hela, MCF7 and HCT116). The choice of secondary antibody is not that critical, you can try any of them.
For more information on antibodies for p53, check following site:
http://p53.free.fr/p53_info/p53_monoclonal_antibodies/p53_Mab.html
As Eugen noted, I would use a mouse monoclonal antibody (the primary antibody) that detects human wild-type p53 protein (because HepG2 cells express a wild-type p53) in immunoblottong. The secondary antibody that is conjugated to HRP (or any other enzyme such as phosphatase) should be able to detect mouse IgG. The dilution and detection conditions need to be optimized.
http://www.genecards.org/cgi-bin/carddisp.pl gene=TP53&search=f1b2452da84551fc91db93425b6a20d9
use this website for p53 gene full details and also it will give all supplier details.
Eugen Werwein and Divaker Choubey were explained very well about p53. So you will use this genecards website for futher details..
Thank you Eugen Werwein, Divaker Choubey and Senthil kumar.
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