Editing Western blot (section)

=== Total protein staining ===
[[File:Western Blot binding.png|thumb|Western blot binding|upright=1.5]]
Total protein staining allows the total protein that has been successfully transferred to the membrane to be visualised, allowing the user to check the uniformity of protein transfer and to perform subsequent normalization of the target protein with the actual protein amount per lane. Normalization with the so-called "loading control" was based on [[immunostaining]] of housekeeping proteins in the classical procedure, but is heading toward total protein staining recently, due to multiple benefits.<ref name="Moritz-2017">{{cite journal | vauthors = Moritz CP | title = Tubulin or Not Tubulin: Heading Toward Total Protein Staining as Loading Control in Western Blots | journal = Proteomics | volume = 17 | issue = 20 | pages = 1600189 | date = October 2017 | pmid = 28941183 | doi = 10.1002/pmic.201600189 | s2cid = 22305461 | url = https://hal.archives-ouvertes.fr/hal-01900776/file/MANSUC~1.PDF }}</ref> At least seven different approaches for total protein staining have been described for western blot normalization: [[Ponceau S]], stain-free techniques, Sypro Ruby, [[Epicocconone]], [[Coomassie brilliant blue|Coomassie R-350]], [[Amido black 10B|Amido Black]], and [[Cyanine|Cy5]].<ref name="Moritz-2017" /> In order to avoid noise of signal, total protein staining should be performed before blocking of the membrane. Nevertheless, post-antibody stainings have been described as well.<ref>{{cite journal | vauthors = Welinder C, Ekblad L | title = Coomassie staining as loading control in Western blot analysis | journal = Journal of Proteome Research | volume = 10 | issue = 3 | pages = 1416–1419 | date = March 2011 | pmid = 21186791 | doi = 10.1021/pr1011476 }}</ref>