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Evaluating dot and Western blots using image analysis and pixel quantification of electronic images

, : Evaluating dot and Western blots using image analysis and pixel quantification of electronic images. Methods in Cell Science 22(4): 313-318

Inexpensive computer imaging technology was used to assess levels of insulin-like growth factor-I (IGF-I) on dot blots (DB) and alpha-Actinin on Western blots (WB). In the first procedure, known IGF-I samples were dotted on nitrocellulose membranes using a vacuum manifold. After the DB were developed and dried, the images were digitized using an HP Deskscan II(R) flat bed scanner, exported into Image-Pro(R) Plus and analyzed by taking the combined mean of 45degree and 135degree sample lines drawn through each dot. Dot blots corresponding to a linear concentration range from 10 to 300 ng IGF-I were assessed by this method. In the second procedure, WB were scanned with a ScanJet 3c(R) flat bed scanner and their backgrounds were clarified using Image-Pro(R) Plus. A second image analysis program, Alpha ImagerTM 2000, was then used to define the boundaries of protein bands, assess pixel number and density, and to obtain final numerical data for quantifying alpha-Actinin on the WB. Collectively, the results of these two studies suggest that specific proteins may be evaluated by using relatively inexpensive image analysis software systems via pixel quantification of electronic images.


PMID: 11549944

DOI: 10.1023/a:1017581302281

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