ImageJ is available for free and will be fine for most applications. Remember that Western blot is only a semiquantitative technique anyway, so I wouldn't get hung up on slight differences between samples and overinterpret results.
If you do need more detailed analysis, stand-alone commercial software is expensive. If you have an imaging system (as opposed to developing on film) somewhere in your University then they always have analysis software as part of the package (e.g. Bio-Rad ImageLab is pretty good and easy to use). You can usually import a 16-bit TIFF file made elsewhere and analyse it, e.g. from scanning a film.
Hope you can find something which works well for your requirements.
ImageJ is available for free and will be fine for most applications. Remember that Western blot is only a semiquantitative technique anyway, so I wouldn't get hung up on slight differences between samples and overinterpret results.
If you do need more detailed analysis, stand-alone commercial software is expensive. If you have an imaging system (as opposed to developing on film) somewhere in your University then they always have analysis software as part of the package (e.g. Bio-Rad ImageLab is pretty good and easy to use). You can usually import a 16-bit TIFF file made elsewhere and analyse it, e.g. from scanning a film.
Hope you can find something which works well for your requirements.
Hi, I agree with Mark, imageJ is a nice (although maybe not always very intuitive) tool, and it's free.
However, keep in mind that any type of film is not linear in its exposure behavior and film is also prone to overexpose. You can maybe get some semi-quantitative information out of film if you have a back-illuminated scanner that can generate 16-bit images, but it will never be truly quantitative. For that, you'd need a CCD camera-based system such as the Fuji/GE LAS-4000 system. With that you achieve more linear exposure characteristics and also have a readout as to whether your signal is saturating the detector (the software would tell you), which you can't tell with film. Out of such a CCD-based system you'd get digital 16-bit TIFF, which is what you need to quantify with imageJ in a reasonable way. The dynamic range of 8-bit images is usually too low to achieve good quantification.
Here some pointers on how to use imagej for densitometry:
http://imagej.nih.gov/ij/docs/guide/146-30.html#infobox:Densitometry
Here another tutorial: http://howtowesternblot.net/data-analysis-3/quantification/
another one: http://lukemiller.org/index.php/2010/11/analyzing-gels-and-western-blots-with-image-j/
Good luck!
See also other threads on Research Gate:
https://www.researchgate.net/post/What_is_the_best_way_to_quantify_protein_fold_change_from_a_western_blot_using_ImageJ
https://www.researchgate.net/post/Quantification_of_protein_band_density_in_X-ray_films_from_ECL_Western_Blots
https://www.researchgate.net/post/How_to_quantify_images_of_western_blot_bands
https://www.researchgate.net/post/Statistics_for_western_blot2
https://www.researchgate.net/post/How_reliable_is_the_chemiluminescent_detection_of_proteins_western_blot_for_relative_quantification
Thank you all for the responses!
I do expect upregulation, but unfortunately not very strong (indeed maybe WB is not the best method in such case, but I do not have choice at the moment).
I started to use infrared detection system (Odyssey) for WB analysis, I believe this will help with quantitative analysis.
Hi Dear A. Pastula
I agree with Dear Mark Thompson. Also you can use of UN-SCAN-IT software but with pay.
Best, Roshdi
- Badges
- Science method