I am using the "ImageJ" software in order to quantify the protein bands on the SDS-PAGE, but I'm not sure it's the best software for this purpose. Does anyone have the experience of using other software to do this? Is there any professional software for SDS-PAGE densitometry?
Usually, the imaging instrument comes with software for densitometry. I have used the Molecular Dynamics software and the Azure Biosystems software. They are pretty similar in all the important respects.
Usually, the imaging instrument comes with software for densitometry. I have used the Molecular Dynamics software and the Azure Biosystems software. They are pretty similar in all the important respects.
Hello, If you do need more detailed analysis, several commercial software available but its expensive. If you have an imaging system they always have analysis software as part of the package.
Bio-Rad Image Lab is good software , You can usually import a 16-bit TIFF file made elsewhere and analyse it, from scanning a film.
Adam B Shapiro , Karen A. Darbinyan & Rasouli H. , thanks for your valuable answers.
Sorry for chiming in so late. Usually, the factor having the greatest influence on the reliability of densitometric SDS-PAGE measurements is not the software package used, but the dynamic range of the staining procedure. For instance, when using Coomassie-stained gels to estimate the purity of purified proteins, it is all too common to overload the gel in an attempt to detect low-abundance contaminants, thereby pushing major species beyond the linear range and underestimating purity (or the opposite situation, loading just enough of major species so that they stay within the linear range but then losing minor species that never make it pass the limit of detection, overestimating purity in the end). There are other factors as well (staining biases depending on protein aminoacid composition; silver is a prominent example but Coomassie suffers from the same issue. Also, band shape plays a role; sometimes the gel section occupied by a major band is wider than the lane so you get better results by "boxing" the entire lane and averaging optical density along the lane than by measuring it along a straight line). And then there is the quality of the densitometer (e.g. using an office document scanner vs a more professional device). Oops sorry for rambling so much. My point is that the software package is low in the list of priorities, Adam's answer is on point, as usual. Using ImageJ is more than fine for most purposes and will make easier reproducing your results. Hope it helps!
Alejandro Martin , Thank you for your valuable advice. Yes, you are right, according to pharmacopoeia methods, gel densitometry commonly used in labs, especially to estimate the percentage of protein of interest and to detect purity of purified proteins. I'm trying to use a professional densitometer instrument, so I hope to minimize errors.
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