9E10 - myc - Western Blotting - HELP!

andrew at cobra.path.monash.edu.au andrew at cobra.path.monash.edu.au
Wed Sep 13 22:13:26 EST 1995


Does anyone know of special comditions for immunblot detection of the 
c-myc epitope using 9E10 monoclonal antibody.

Currently I transfer my PAGE gel to nitrocellulose,
block with 5% skim milk powder in PBS,
probe with 9E10 supernatant,
wash with PBS/Tween
probe with antimouse-HRPO in 5% skim milk powder in PBS,
wash with PBS/Tween, then detect with ECL.

However, I am getting many non-specific bands coming up, and I thought 
that 9E10 was quite specific for myc.

Does anyone know of a way to elminate these background bands?

Thanks in advance,

Andrew (Melbourne, Australia).





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