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densitometry Analysis - (Nov/27/2013 )

hi

which instrument are you using for densitometry analysis ?

and , what are the common pitfalls observed for densitometry analysis

-rick112-

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.

-bob1-

hi

thanks bob1

what about when used with SDS-PAGE coomassie staining  especially with respect to background and artifacts ???

-rick112-

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.

-bob1-

we are evaluating both Imagescanner from GE and GS900 from Biorad

anyone has experience of one of these instruments ??

-rick112-