densitometry Analysis - (Nov/27/2013 )
which instrument are you using for densitometry analysis ?
and , what are the common pitfalls observed for densitometry analysis
A decent scanner and ImageJ or Quantity1 software (proprietary).
Pitfalls - exposure is critical, any pixels that are saturated will throw off the analysis. Technique is not easily used across blots. Standard AP/HRP chemistry is not very linear in response, ideally use fluorescent antibodies.
what about when used with SDS-PAGE coomassie staining especially with respect to background and artifacts ???
Artifacts will always throw off an analysis - you need nice clean gels with good separation of sharp bands - run big gels of the appropriate percentage, none of these crappy tiny Biorad gels. Background subtraction is a must.
we are evaluating both Imagescanner from GE and GS900 from Biorad
anyone has experience of one of these instruments ??