Housekeeping proteins: How useful are they in skeletal muscle diabetes studies and muscle hypertrophy models?

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....

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Bibliographic Details
Published in:Analytical biochemistry 2016-07, Vol.504, p.38-40
Main Authors: Fortes, Marco Aurélio Salomão, Marzuca-Nassr, Gabriel Nasri, Vitzel, Kaio Fernando, da Justa Pinheiro, Carlos Hermano, Newsholme, Philip, Curi, Rui
Format: Article
Language:English
Subjects:
Actinin - analysis
Actins - analysis
Animals
Blotting, Western
Diabetes Mellitus, Experimental - chemically induced
Diabetes Mellitus, Experimental - metabolism
Glyceraldehyde-3-phosphate dehydrogenase
Glyceraldehyde-3-Phosphate Dehydrogenases - analysis
Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism
Hyperglycemia - chemically induced
Hyperglycemia - metabolism
Male
Muscle, Skeletal - chemistry
Muscle, Skeletal - metabolism
Ponceau S
Rats
Rats, Wistar
Streptozocin
Tubulin - analysis
α-Actinin
α-Tubulin
β-Actin
γ-Tubulin
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Summary: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.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2016.03.023