Hello,
I am trying to look for the expression on IL-6 in human Prostate Cancer cell lines. I was wondering wich antibody should I use to see the level of expression of IL-6 protein in the LNcap cell lines, by Western blot.
I am kind of confused, since this is a protein with lot of choices available.Any help is appreciated.
Thanks.
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#2
protolder
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Posted 28 September 2010 - 09:34 PM
Hola, the suppliers of antibodies have all the necesary information about aplications, so first you have to choice anti human Il6 for WB.Monoclonal is more specific, source conditioned the secondary that you have to use, mouse is commun, and inmunogen; the ab has been prepared with an specific peptide or with the complete protein. Dilution of use, it is described in the data sheet.It could be that the ab recognizes IL6 from other species, as monkey rat etc, but for your application it doesnŽt matter.The sole remaining question is the price, but I supose that there areŽnt big differences between them . So, if I were you I would choice a monoclonal mouse anti human IL6 for WB if it were possible against all the molecule. and for secundary goat anti mouse HRP or with fluorescence marker. See the Santa Cruz catalogue written or on line. There are images aof the results of WB with each antibody. Good luck
#3
friend26
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Posted 29 September 2010 - 09:41 AM
protolder, on 28 September 2010 - 09:34 PM, said:
Hola, the suppliers of antibodies have all the necesary information about aplications, so first you have to choice anti human Il6 for WB.Monoclonal is more specific, source conditioned the secondary that you have to use, mouse is commun, and inmunogen; the ab has been prepared with an specific peptide or with the complete protein. Dilution of use, it is described in the data sheet.It could be that the ab recognizes IL6 from other species, as monkey rat etc, but for your application it doesnŽt matter.The sole remaining question is the price, but I supose that there areŽnt big differences between them . So, if I were you I would choice a monoclonal mouse anti human IL6 for WB if it were possible against all the molecule. and for secundary goat anti mouse HRP or with fluorescence marker. See the Santa Cruz catalogue written or on line. There are images aof the results of WB with each antibody. Good luck
Thanks a lot for the information. It is really helpful. I will try to use as per your suggestions. I hope it should work, since I am using human Prostate cancer cell lines.
