Heating induces aggregation and decreases detection of serotonin transporter protein on western blots.
In recent years, there has been growing interest in the use of Western blot analysis to monitor changes in the abundance of the serotonin transporter (SERT) protein. In the Western blot procedure, heat denaturation is a common, early step. We now report that heating samples to 90 degrees C decreases the abundance of the SERT protein band and causes dispersion of a majority of the SERT signal to a high molecular weight smear. These observations are in keeping with the fact that heating can influence the electrophoretic behavior of some proteins. By omitting the heat denaturation step in the Western blot procedure, better detection of the SERT protein is achieved.
McLane
Michael W
MW
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21224, USA.
Hatzidimitriou
George
G
Yuan
Jie
J
McCann
Una
U
Ricaurte
George
G
D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D006361:Heating / Q000009:adverse effects; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000494:pharmacology; D011489:Protein Denaturation / Q000528:radiation effects; D051381:Rats; D018490:Serotonin Agents / Q000494:pharmacology; D050486:Serotonin Plasma Membrane Transport Proteins / Q000378:metabolism*
D018490:Serotonin Agents; D050486:Serotonin Plasma Membrane Transport Proteins; D018817:N-Methyl-3,4-methylenedioxyamphetamine