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Yesterday, I used your p19ARF antibody(ab80) for my samples. Also I set NIH3T3 whole cell lysate as a positive control, but it didn’t work. (no band.)
I borrowed this cell line from other lab. After I cultured, I extracted lysate by myself.
I think there is some possibility that my NIH3T3 had any problems. So, I plan to order your cell lysate and check some mouse embryonic fibroblast cell line; ab7179,ab14874,ab7901,ab3874.
I’d like to ask you which is the best as positive control for p19ARF antibody? Or is there no difference among them?
Asked on Oct 11 2012
Thank you very much for contacting us with your enquiry.
I'm sorry to hear that there were no bands in Western blot using this antibody. I would expect any of the MEF cell lines to work as a positive control, but I suggest using a whole cell lysate (ab14874 is a nuclear fraction) since that is what we have tested with ab80. What kind of samples are you using other than the MEF lysate?
If you'd like to send some more details about your protocol ( how much protein was loaded, dilution of antibody, blocking solution, etc) I'd be happy to take a look and see if we have any suggestions to improve the results.
I look forward to hearing from you. If there's anything else that we can do for you, please let me know and I'll be happy to help.
Answered on Oct 11 2012