Ghost bands on Western blot?

Darren Fast fast at biochem.wisc.edu
Mon Mar 27 14:27:18 EST 1995


In article <3l6i2k$pr6 at hippo.ru.ac.za>, g88d4915 at hippo.ru.ac.za (Mr TG
Downing) wrote:

> Can anyone shed any light on the following: The protein we're looking at
> appears as ghost bands while non-specific stuff comes out nicely. Using
> commercial monoclonal (which is OK according to supplier) and a second
> Ap-conjugate afterwards. Someone told me that this may be due to
> "masking" by the primary antibody and "outcompetition" for the secondary
> by non-specific contaminants. Any other suggestions/ideas?
> TIA

I've seen this before when my antibody concentrations were too high and
when I've loaded too much of my protein onto the gel.

Darren

-- 
Darren Fast
Dept of Biochemistry
University of Wisconsin-Madison
ph. (608)-262-4705
fax (608)-262-3453
e-mail FAST at biochem.wisc.edu



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