| Rabbit Anti-phospho-ERK1 (Thr202/Tyr204) + ERK2 (Thr183/Tyr185) antibody |
| 反应物种（预测） |
Chicken,Dog,Cow,Horse,Rabbit,GuineaPig |
| 产品应用（已验证） |
IHC,FCM |
| 产品应用（可尝试） |
WB,IF,ELISA |
| 推荐稀释比例 |
WB=1:500-2000,Elisa=1:5000-10000,IHC-P=1:100-500,IHC-F=1:100-500,Flow Cyt=1μg /test,IF=1:100-500, |
| 研究领域 |
神经生物学,信号转导,干细胞,激酶和磷酸酶 |
| 标签 |
Array |
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Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-ERK1 (Thr202Tyr204) + ERK2 (Thr183Tyr185)) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Blank control: MCF7.
Primary Antibody (green line): Rabbit Anti-Phospho-ERK1 (Thr202/Tyr204) + ERK2 (Thr183/Tyr185) antibody (bs-1646R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Sample:
Heart (Mouse) Lysate at 30 ug
SP2/0 Cell Lysate at 30 ug
Primary: Anti-phospho-ERK1(Thr202/Tyr204)+ERK2(Thr183/Tyr185) (bs-1646R) at 1:200 dilution;
Secondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 3000 dilution;
Predicted band size : 42kD
Observed band size : 42kD
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Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (P-ERK 1+ ERK 2) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
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From 《Cancer Medicine》(2016.6): PublitionDirect effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes , IF:2.5
Author: Noriyoshi Iriyama, Yoshihiro Hatta & Masami Takei
Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, Tokyo, Japan
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Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-phospho-ERK1(Thr202/Tyr204)+ERK2(Thr183/Tyr185) Polyclonal Antibody, Unconjugated(bs-1646R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Paraformaldehyde-fixed, paraffin embedded (rat lung tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (P-ERK 1+ ERK 2) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
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Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-ERK1 (Thr202Tyr204) + ERK2 (Thr183Tyr185)) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-ERK1 (Thr202Tyr204) + ERK2 (Thr183Tyr185)) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-ERK1 (Thr202Tyr204) + ERK2 (Thr183Tyr185)) Polyclonal Antibody, Unconjugated (bs-1646R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
RRID：AB_10855892
产品名称：Rabbit Anti-phospho-ERK1 (Thr202/Tyr204) + ERK2 (Thr183/Tyr185) antibody
别名： Erk1 (pT202/pY204) + Erk2 (pT185/pY187); p44/42 MAPK (Erk1/2) (Thr202/Tyr204); Erk1(pT202/pY204)+Erk2(pT185/pY187); ERK 1; ERK 2; ERK-2; ERK1; ERK2; ERT1; ERT2; Extracellular signal regulated kinase 1; Extracellular signal regulated kinase 1; Extracellula
中文名称：磷酸化丝裂原活化蛋白激酶1/2抗体
英文名称：Rabbit Anti-phospho-ERK1 (Thr202/Tyr204) + ERK2 (Thr183/Tyr185) antibody
抗体来源： Rabbit
克隆类型：多克隆
细胞定位：细胞核,细胞浆
性 状：Liquid
亚 型：IgG
纯化方法：affinity purified by Protein A
保存条件：Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原：KLH conjugated Synthesised phosphopeptide derived from mouse ERK1/2 around the phosphorylation site of Thr202/Tyr204
抗原表位：FL(p-T)E(p-Y)VA
SWISS：P28482
Gene ID ：5595
Human Gene ID：5595
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. The activation of this kinase requires its phosphorylation by upstream kinases. Upon activation, this kinase translocates to the nucleus of the stimulated cells, where it phosphorylates nuclear targets. Two alternatively spliced transcript variants encoding the same protein, but differing in the UTRs, have been reported for this gene.
Function：Serine/threonine kinase which acts as an essentialcomponent of the MAP kinase signal transduction pathway. MAPK1/ERK2and MAPK3/ERK1 are the 2 MAPKs which play an important role in theMAPK/ERK cascade. They participate also in a signaling cascadeinitiated
Subunit：Binds both upstream activators and downstream substratesin multimolecular complexes. Interacts with ADAM15, ARHGEF2, ARRB2,DAPK1 (via death domain), HSF4, IER3, IPO7, DUSP6, NISCH, SGK1, andisoform 1 of NEK2. Interacts (via phosphorylated form) with TPR(v
Subcellular Location：Cytoplasm, cytoskeleton, spindle (Bysimilarity). Nucleus. Cytoplasm, cytoskeleton, centrosome (Bysimilarity). Cytoplasm. Note=Associated with the spindle duringprometaphase and metaphase (By similarity). PEA15-binding andphosphorylated DAPK1 promote its c
Tissue Specificity：Widely expressed.
Post-translational modifications：Dually phosphorylated on Thr-183 and Tyr-185, which activatesthe enzyme. Ligand-activated ALK induces tyrosine phosphorylation(By similarity). Dephosphorylated by PTPRJ at Tyr-185 (Bysimilarity). Phosphorylated upon FLT3 and KIT signaling (Bysimilarity).
Similarity：Dually phosphorylated on Thr-183 and Tyr-185, which activatesthe enzyme. Ligand-activated ALK induces tyrosine phosphorylation(By similarity). Dephosphorylated by PTPRJ at Tyr-185 (Bysimilarity). Phosphorylated upon FLT3 and KIT signaling (Bysimilarity).
Important Note：This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
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