handling of cmyc antibody

christian christian.troidlNOchSPAM at kerckhoff.mpg.de.invalid
Thu Jun 8 09:04:54 EST 2000


Hello colleagues!
I have problems with the detection of cmyc fusion proteins
(cmyc fused to human myosin binding protein C) recruted from
transfected COS cells or neonatal cardiocytes.
The monoclonal antibody is delivered by Boehringer Mannheim.
The dilution I used was 1:250 in 1% blocking buffer (milk
powder).
I tried to incubate with the antibody for two hours and over
nigth. Once I got a signal with crude extracts of COS cells
but only with a high amount of total protein. I can´t
reproduce this result, I always get no significant signal.
Please give me some hints how to use this antibody or how to
improve my Western blot technique.



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