Normalization of Western blot data is an analytical step that is performed to compare the relative abundance of a specific protein across the lanes of a blot or gel under diverse experimental treatments, or across tissues or developmental stages.[1][2] The overall goal of normalization is to minimize effects arising from variations in experimental errors, such as inconsistent sample preparation, unequal sample loading across gel lanes, or uneven protein transfer, which can compromise the conclusions that can be obtained from Western blot data.[1] Currently, there are two methods for normalizing Western blot data: (i) housekeeping protein normalization and (ii) total protein normalization.[1][2][3][4]
^ abcTaylor, SC; Berkelman, T; Yadav, G; Hammond, M (2016-08-23). "A defined methodology for reliable quantification of Western blot data". Mol. Biotechnol. 55 (3): 217–26. doi:10.1007/s12033-013-9672-6. PMC 3840294. PMID 23709336.
^ abThellin, O.; Zorzi, W.; Lakaye, B.; De Borman, B.; Coumans, B.; Hennen, G.; Grisar, T.; Igout, A.; Heinen, E. (1999-10-08). "Housekeeping genes as internal standards: use and limits". Journal of Biotechnology. 75 (2–3): 291–295. doi:10.1016/s0168-1656(99)00163-7. hdl:2268/3661. ISSN 0168-1656. PMID 10617337.
^Aldridge, Georgina M.; Podrebarac, David M.; Greenough, William T.; Weiler, Ivan Jeanne (2008-07-30). "The use of total protein stains as loading controls: An alternative to high-abundance single-protein controls in semi-quantitative immunoblotting". Journal of Neuroscience Methods. 172 (2): 250–254. doi:10.1016/j.jneumeth.2008.05.003. PMC 2567873. PMID 18571732.
^Collins, Mahlon A.; An, Jiyan; Peller, Danielle; Bowser, Robert (2015-08-15). "Total protein is an effective loading control for cerebrospinal fluid western blots". Journal of Neuroscience Methods. 251: 72–82. doi:10.1016/j.jneumeth.2015.05.011. PMC 4540354. PMID 26004848.
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