Peptide Competition. Recombinant eIF2Bepsilon treated with lambda phosphatase (1) or GSK-3beta (2-5) were added to background extracts and resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the eIF2Bepsilon [pS539] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphoserine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab and quote;)2 anti-rabbit IgG alkaline phosphatase (Cat. no. ALI4405) and signals were detected using the Tropix WesternStar™ method. The data show that only the peptide corresponding to eIF2Bepsilon [pS539] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific. The recombinant eIF2Bepsilon was kindly provided by Dr. Scot R. Kimball, Penn State University.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Tag, Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of rat eIF2Be that contains serine 535 (corresponding to serine 539 in the human sequence).|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 6 publications below|
This gene encodes one of five subunits of eukaryotic translation initiation factor 2B (EIF2B), a GTP exchange factor for eukaryotic initiation factor 2 and an essential regulator for protein synthesis. Mutations in this gene and the genes encoding other EIF2B subunits have been associated with leukoencephalopathy with vanishing white matter.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Pharmacological inhibition of GSK-3 in a guinea pig model of LPS-induced pulmonary inflammation: II. Effects on skeletal muscle atrophy.
44-530G was used in western blot to use a guinea pig model of LPS-induced pulmonary inflammation to elucidate the pharmacological inhibition of GSK-3
|Verhees KJ,Pansters NA,Baarsma HA,Remels AH,Haegens A,de Theije CC,Schols AM,Gosens R,Langen RC||Respiratory research (14:null)||2013|
|Glycogen synthase kinase-3ß is required for the induction of skeletal muscle atrophy.||Verhees KJ,Schols AM,Kelders MC,Op den Kamp CM,van der Velden JL,Langen RC||American journal of physiology. Cell physiology (301:C995)||2011|
|Human||Not Cited||Nutrient provision increases signalling and protein synthesis in human skeletal muscle after repeated sprints.||Coffey VG,Moore DR,Burd NA,Rerecich T,Stellingwerff T,Garnham AP,Phillips SM,Hawley JA||European journal of applied physiology (111:1473)||2011|
|Rat||1:2000||Ectopic expression of eIF2Bepsilon in rat skeletal muscle rescues the sepsis-induced reduction in guanine nucleotide exchange activity and protein synthesis.||Tuckow AP,Vary TC,Kimball SR,Jefferson LS||American journal of physiology. Endocrinology and metabolism (299:E241)||2010|
|Human||Not Cited||Pulmonary artery smooth muscle hypertrophy: roles of glycogen synthase kinase-3beta and p70 ribosomal S6 kinase.||Deng H,Hershenson MB,Lei J,Anyanwu AC,Pinsky DJ,Bentley JK||American journal of physiology. Lung cellular and molecular physiology (298:L793)||2010|
|Rat||Not Cited||Resistance exercise increases muscle protein synthesis and translation of eukaryotic initiation factor 2Bepsilon mRNA in a mammalian target of rapamycin-dependent manner.||Kubica N,Bolster DR,Farrell PA,Kimball SR,Jefferson LS||The Journal of biological chemistry (280:7570)||2005|