Dear all, after creating several mutants containing a myc-tag, I have discovered that in infection conditions, my anti-myc antibodies are not able to recognize the tag but they do recognize it in blots where the bacteria lysates are loaded alone. I am sure that the reason is that the amount of bacteria bound to the cells is only 1/10th of the lysate. Still, I need to see the myc in the infections. I have used the Cell Signaling mouse anti-myc (very good ) and the THE(TM) anti-myc from Genscript (not that good, but it works!). Both are monoclonal anti-mouse.
Any input will be appreciated. I am not keen in doing a new tag-mutants (6 to 8 months work!)
Thanks
Hi Krzysztof,
thanks for the answer. about selfmade... I have no rabbit...
And about the infection and degradation of the proteins: I can detect the full protein with another antibody (a selfmade!), but the tag (that is in the middle of teh protein) I cannot. Additionally, my infections are from 5 min to 4 hours... and in all I can see the protein with the other antibody... but not the tag... I will try to see if I can find a polyclonal somewhere... wish me luck!... and thanks again!
Hi,
we are using the 9E10 (e.g. from Sigma). It's a monoclonal murine antibody. The antibody has a good affinity and detection limit.
Michael
c-myc antibody (9E10): sc-40 from santa cruz work very well for us (IP and WB)
Thanks for all your answers... I will be doing the tests soon...
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