[Protein-analysis] "ghost" dots on blot?
(by jessica_krakow from targetdiscovery.com)
Tue Jul 17 16:35:53 EST 2007
What causes negative or "ghost" bands/spots on a western or dot blot?
I did a dot blot for determining what dilutions of primary and
secondary Abs to use. I spotted about 10 micrograms of total protein
onto nitrocellulose for each spot and probed with various dilutions
of my primary and secondary Abs.
I got no signal at all in every spot. The area where the protein sat
was much lighter than background. Too much protein? Too much Ab?
Thanks in advance for your help!
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