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densitometry Analysis


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4 replies to this topic

#1 rick112

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Posted 27 November 2013 - 10:20 PM

hi

which instrument are you using for densitometry analysis ?

and , what are the common pitfalls observed for densitometry analysis



#2 bob1

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Posted 28 November 2013 - 12:04 AM

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.



#3 rick112

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Posted 28 November 2013 - 12:53 AM

hi

thanks bob1

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



#4 bob1

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Posted 28 November 2013 - 12:40 PM

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.



#5 rick112

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Posted 29 November 2013 - 12:50 AM

we are evaluating both Imagescanner from GE and GS900 from Biorad

anyone has experience of one of these instruments ??






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