title: Differential detection of phosphorylated glycogen synthase kinase 3 alpha and beta depending on blocking conditions.
abstract: It is generally recommended that immunoblots using phosphospecific antibodies be performed using bovine serum albumin (BSA) instead of milk. There are two subtypes of glycogen synthase kinase 3 (GSK3), GSK3 alpha and GSK3 beta, with apparent molecular weights of 51 and 49 kDa, respectively. Here we show that immunoblots performed using 5% milk allow the detection of both phosphorylated GSK3 alpha and phosphorylated GSK3 beta, whereas only phosphorylated GSK3 alpha is visible when immunoblots are performed using 3% BSA.
authors.affiliation: Institute of Cell Signalling, School of Biomedical Sciences, University of Nottingham, Nottingham NG7 2UH, UK.
authors.forename: M L
authors.initials: ML
authors.lastname: Latif
authors.forename: T S
authors.initials: TS
authors.lastname: Pillay
mesh_terms: D000818:Animals; D016466:CHO Cells; D002417:Cattle; D006224:Cricetinae; D003412:Cricetulus; D004789:Enzyme Activation / Q000187:drug effects; D038362:Glycogen Synthase Kinase 3 / Q000032:analysis* / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007328:Insulin / Q000494:pharmacology; D010766:Phosphorylation; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D007328:Insulin; D011972:Receptor, Insulin; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3; C448021:glycogen synthase kinase 3 alpha