Usually in western blot there is a single band for a target protein and we can measure its density and calculate with reference to an internal control for normalization. the problem occurs when there are many specific bands are detected in western blot for a target protein, eg. collagens.
How shall I measure the bands density?
Also, please, I appreciate any examples in published papers.
Thank you.
You can analyze band intensity using ImageJ software. Using imageJ you can select the bands of your interest from the blot.
Hi there,
You may quantify every band corresponding to your protein as each band is specific of one specific state. So you may qualitatively and quantitatively analyze your blot.
Thanks Dominique Liger, I also think that analysis of bands one by one is the best way, but I cam not find similar results in papers, thanks for reply.
I must sadly and repeatedly say that notorious Western blotting is not applicable to hydrophobic proteins (please see file; IEF for hydrophobic proteins).
Instead of notorious Western blot, I would like to recommend to use 1stly to separate the proteins by HPLC-Surf-SEC method (please see file; SEC column 300 A silica).
After the fractionation, you should use 2ndly the PDMD (protein-direct-microsequencing-deciphering) method to identify the protein (please see file; HepG2 fucoidan). These 2 HPLC methods are quantitative.
This is not a my propaganda at all, but this recommendation is only saying the holy and/or sacred truth.
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