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β‐Actin is not a reliable loading control in Western blot analysis
Author(s) -
Dittmer Angela,
Dittmer Jürgen
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500785
Subject(s) - western blot , actin , antibody , incubation , dot blot , blot , biology , microbiology and biotechnology , chemistry , biochemistry , immunology , gene
β‐Actin is often used as a loading control in Western blot analysis. We analyzed the ability of β‐actin‐specific antibodies to recognize differences in protein loading. We found that, at higher total protein loads as required for the detection of low‐abundance proteins, β‐actin‐specific antibodies failed to distinguish differences in actin protein levels. Diluting the antibody working solution or changing the incubation time had little effect on this phenomenon. This shows that β‐Actin is not a reliable loading control in Western blot analysis. In general, it appeared that, at longer incubation times, antibodies seem to be less able to pick up differences in the level of its target protein.