Application
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)2 anti-rabbit IgG alkaline phosphatase and signals were detected. 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| Product Name | EIF2B epsilon (phospho Ser539) antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal to eIF2B epsilon ( phospho S539 )Eukaryotic Initiation Factor 2B (eIF-2B) plays a key role by acting as a guanine nucleotide exchange factor in the initiation of mRNA translation in eukaryotic cells. The activity of eIF-2B is regulated by a wide array of conditions. Stress inhibits its action, while stimulation of protein synthesis results in activation. eIF-2B epsilon, an 82 kDa subunit of eIF-2B, is phosphorylated at serine 539 by glycogen synthase kinase 3 (GSK 3), which leads to inhibition of its guanine nucleotide exchange activity. |
| Immunogen | Synthetic peptide (Rat) derived from the region of rat eIF-2B epsilon that contains serine 535 (corresponding to serine 539 in the human sequence). |
| Modification | p-Ser539 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | WB |
| WB: Use at a concentration of 0.1 - 1.0 μg/ml. Predicted molecular weight: 82 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | EIF-2B 8893 EIF2Bepsilon EIF2B epsilon EIF2B5 CACH Q13144 LVWM CLE EIF2B 603896 EIF 2B EIF2B epsilon (phospho S539) EIF2B epsilon (phospho Ser539) EIF2B epsilon (pS539) EIF2B epsilon (pSer539) EIF2B epsilon phospho S539 EIF2B epsilon phos |
|---|---|
| Target | EIF2B epsilon |
Application
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)2 anti-rabbit IgG alkaline phosphatase and signals were detected. 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| Positive Control | Recombinant eIF-2B epsilon treated with GSK 3 beta and lambda phosphatase, CHO-T cells transfected with human insulin receptor (IR) +/- insulin |
|---|---|
| Application Notes | WB: Use at a concentration of 0.1 - 1.0 μg/ml. Predicted molecular weight: 82 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
| Form | Liquid |
|---|---|
| Storage Instructions | Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | Preservative: 0.05% Sodium Azide; Constituents: PBS, 50% glycerol, 1.0mg/ml BSA (IgG, protease free). pH 7.3 |
