Application
Extracts of HEK293 cells transiently transfected with plasmids expressing ERK5 kinase domain (ERK5kin) and constitutively activated MEK5D-D were 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 ℃, then incubated with the ERK5 (phospho Thr218) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphothreonine-containing peptide (3), a generic phosphotyrosine-containing peptide (4), the phosphopeptide derived from the corresponding region of ERK1&2 (5), or the phosphopeptide immunogen (6). The data show that while there is some cross-reactivity with ERK1&2, only the phosphopeptide corresponding to ERK5 [pTpY218/220] completely blocks the antibody signal, demonstrating the specificity of the antibody. NOTE: The antibody signal appears at ~50 kDa as this is the molecular weight of the transiently transfected ERK5 kinase domain.| Product Name | ERK5 (phospho Thr218/Tyr220) antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit Polyclonal antibody to ERK5 (phospho Thr218/Tyr220)Extracellular signal-related protein kinase 5/big mitogen-activated protein kinase 1 (ERK5/BMK1) is an 88 kDa member of the MAP kinase superfamily that becomes fully activated when phosphorylated on threonine 218 and tyrosine 220 by MEK5. ERK5 is activated in response to certain growth factors and bioactive peptides, however the upstream activators are distinct from those that activate ERK1&2. |
| Immunogen | Synthetic phosphopeptide derived from the region of human ERK5 that contains threonine 218 and tyrosine 220. |
| Modification | p-Thr218/Tyr220 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | IHC-FWB |
| IHC-Fr: Use at an assay dependent dilution. WB: Use at a dilution of 1/1000. Predicted molecular weight: 88 kDa. Due to the low abundance and low levels of activation of endogenous ERK5, over-expression or immunoprecipitation of ERK5 protein may be required to detect activation. Optimal dilutions/concentrations should be determined by the end user.: |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml (Please refer to the vial label for the specific concentration) |
| Purification | Affinity purified |
| Gene Synonyms | MAPK7 602521 ERK5 ERK4 PRKM7 BMK1 5598 Q13164 ERK5 (phospho T218) ERK5 (phospho Thr218) ERK5 (pT218) ERK5 (pThr218) ERK5 phospho T218 ERK5 phospho Thr218 Phospho ERK5 (T218) Phospho ERK5 (Thr218) Phospho ERK5 (pT218) Phospho ERK5 (pThr218) |
|---|---|
| Target | ERK5 (phospho Thr218/Tyr220) |
| Tissue Specificity | Some cross-reactivity is observed with endogenous ERK1 and 2 (44 and 42 kDa, respectively) due to the high levels of expression and activation of this protein typically observed with most cell types. |
Application
Extracts of HEK293 cells transiently transfected with plasmids expressing ERK5 kinase domain (ERK5kin) and constitutively activated MEK5D-D were 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 ℃, then incubated with the ERK5 (phospho Thr218) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphothreonine-containing peptide (3), a generic phosphotyrosine-containing peptide (4), the phosphopeptide derived from the corresponding region of ERK1&2 (5), or the phosphopeptide immunogen (6). The data show that while there is some cross-reactivity with ERK1&2, only the phosphopeptide corresponding to ERK5 [pTpY218/220] completely blocks the antibody signal, demonstrating the specificity of the antibody. NOTE: The antibody signal appears at ~50 kDa as this is the molecular weight of the transiently transfected ERK5 kinase domain.| Positive Control | HEK293 cells transiently co-transfected with plasmids expressing ERK5 kinase domain (ERK5kin)and constitutively activated MEK5 (MEK5D-D) |
|---|---|
| Application Notes | IHC-Fr: Use at an assay dependent dilution. WB: Use at a dilution of 1/1000. Predicted molecular weight: 88 kDa. Due to the low abundance and low levels of activation of endogenous ERK5, over-expression or immunoprecipitation of ERK5 protein may be required to detect activation. Optimal dilutions/concentrations should be determined by the end user.: |
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution. Store at 4ºC short term. For extended storage aliquot and store at -20ºC or below. Avoid freeze-thaw cycles. |
| Storage Buffer | Preservative: 0.05% Sodium Azide; Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3 |
