WB
Western blot analysis of Phospho-Erk1 (T202)+Erk2 (T185) on different lysates with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: Jurkat treated with 200ng/mL PMA for 35 minutes cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate, Lane 5: C6 cell lysate, Lane 6: C6 treated with 200nM PMA for 30 minutes cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
ICC staining Phospho-Erk1(T202)+Erk2(T185) in NIH/3T3 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.ICC/IF
ICC staining Phospho-Erk1(T202)+Erk2(T185) in MCF-7 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.FC
Flow cytometric analysis of Phospho-Erk1 (T202)+Erk2 (T185) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Phospho-Erk1 (T202)+Erk2 (T185) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines, and research investigators consider it an important target in the diagnosis and treatment of cancer. Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway. MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK and the transcription factor Elk-1. p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs, along with MEK inhibitors, such as U0126 and PD98059. |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Thr185 of Human Erk2. |
| Modification | p-T202+T185 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICCIHCWB |
| WB:1:1,000 ICC:1:100-1:500 IHC:1:50-1:200 FC:1:50-1:100 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Molecular Weight(MW) | 42/44 kDa |
|---|---|
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of Phospho-Erk1 (T202)+Erk2 (T185) on different lysates with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. Lane 1: Jurkat cell lysate, Lane 2: Jurkat treated with 200ng/mL PMA for 35 minutes cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate, Lane 5: C6 cell lysate, Lane 6: C6 treated with 200nM PMA for 30 minutes cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
ICC staining Phospho-Erk1(T202)+Erk2(T185) in NIH/3T3 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC/IF
ICC staining Phospho-Erk1(T202)+Erk2(T185) in MCF-7 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-Erk1 (T202)+Erk2 (T185) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
FC
Flow cytometric analysis of Phospho-Erk1 (T202)+Erk2 (T185) was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Positive Control | A549, NIH/3T3, MCF-7, human lung cancer tissue, human kidney tissue. |
|---|---|
| Application Notes | WB:1:1,000 ICC:1:100-1:500 IHC:1:50-1:200 FC:1:50-1:100 |
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
