title: Housekeeping proteins: How useful are they in skeletal muscle diabetes studies and muscle hypertrophy models?
abstract: The use of Western blot analysis is of great importance in research, and the measurement of housekeeping proteins is commonly used for loading controls. However, Ponceau S staining has been shown to be an alternative to analysis of housekeeping protein levels as loading controls in some conditions. In the current study, housekeeping protein levels were measured in skeletal muscle hypertrophy and streptozotocin-induced diabetes experimental models. The following housekeeping proteins were investigated: glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin, α-tubulin, γ-tubulin, and α-actinin. Evidence is presented that Ponceau S is more reliable than housekeeping protein levels for specific protein quantifications in Western blot analysis.
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Butantã, São Paulo 05508-900, Brazil. Electronic address: markofortes@hotmail.com.
authors.forename: Marco Aurélio Salomão
authors.initials: MA
authors.lastname: Fortes
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Butantã, São Paulo 05508-900, Brazil.
authors.forename: Gabriel Nasri
authors.initials: GN
authors.lastname: Marzuca-Nassr
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Butantã, São Paulo 05508-900, Brazil.
authors.forename: Kaio Fernando
authors.initials: KF
authors.lastname: Vitzel
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Butantã, São Paulo 05508-900, Brazil.
authors.forename: Carlos Hermano
authors.initials: CH
authors.lastname: da Justa Pinheiro
authors.affiliation: School of Biomedical Sciences, Curtin Health Innovation Research Institute (CHIRI) Biosciences, Curtin University, Bentley, Perth, Western Australia 6102, Australia.
authors.forename: Philip
authors.initials: P
authors.lastname: Newsholme
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, Butantã, São Paulo 05508-900, Brazil.
authors.forename: Rui
authors.initials: R
authors.lastname: Curi
mesh_terms: D000185:Actinin / Q000032:analysis*; D000199:Actins / Q000032:analysis*; D000818:Animals; D015153:Blotting, Western; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis* / Q000378:metabolism; D006943:Hyperglycemia / Q000139:chemically induced / Q000378:metabolism; D008297:Male; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013311:Streptozocin; D014404:Tubulin / Q000032:analysis*
chemical_list: D000199:Actins; D014404:Tubulin; D000185:Actinin; D013311:Streptozocin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Glyceraldehyde-3-phosphate dehydrogenase; Ponceau S; α-Actinin; α-Tubulin; β-Actin; γ-Tubulin