Tissue/cell: rat brain tissue; 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) Polyclonal Antibody, Unconjugated(bs-1645R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
- ￥1180 - 2800
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- bs-1645R
- 2025年10月24日
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50ul/100ul/200ul
| 规格： | 50ul | 产品价格： | ￥1180.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥1980.0 |
| 规格： | 200ul | 产品价格： | ￥2800.0 |
| 产品编号 | bs-1645R |
| 英文名称 | phospho-Erk1 (Thr202 + Tyr204) Rabbit pAb |
| 中文名称 | 磷酸化丝裂原活化蛋白激酶1抗体 |
| 英文别名 | Erk1(pT202/pY204); ERK/MAPK(phospho T202/Y204); ERK1(phospho T202); p-ERK1(phospho T202); p44/42 MAP Kinase(phospho-Thr202); ERK; ERK-1; ERT 2; ERT2; Extracellular Signal Regulated Kinase 1; Extracellular signal related kinase 1; Extracellular signal-regulated kinase 1; HGNC6877; HS44KDAP; HUMKER1A; Insulin Stimulated MAP2 Kinase; Insulin-stimulated MAP2 kinase; MAP kinase 1; MAP kinase 3; MAP Kinase; MAP kinase isoform p44; MAPK 1; MAPK 3; MAPK; MAPK1; Mapk3; MGC20180; Microtubule Associated Protein 2 Kinase; Microtubule-associated protein 2 kinase; Mitogen Activated Protein Kinase 3; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 3; MK03_HUMAN; OTTHUMP00000174538; OTTHUMP00000174541; p44 ERK1; p44 MAPK; p44-ERK1; p44-MAPK; P44ERK1; P44MAPK; PRKM 3; PRKM3; Protein Kinase Mitogen Activated 3. |
| 产品应用 | IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat (Chicken, Dog, Cow, Horse, Rabbit, GuineaPig) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from rat ERK1 around the phosphorylation site of Thr201/204 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 43 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Neuroscience > Neurology process > Neurodegenerative disease > Alzheimer's disease > Tangles & Tau Signal Transduction > Protein Phosphorylation > Ser / Thr Kinases > MAPK Pathway Stem Cells > Signaling Pathways > TGF beta > Cytoplasmic |
| 亚基 | 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(via C-terminus region and phosphorylated form); the interactionrequires dimerization of MAPK1/ERK2 and increases following EGFstimulation. Interacts (phosphorylated form) withCAV2 ('Tyr-19'-phosphorylated form); the interaction, promoted byinsulin, leads to nuclear location and MAPK1 activation. Interacts with DCC. Interacts withMORG1, PEA15 and MKNK2. MKNK2 isoform 1 binding prevents fromdephosphorylation and inactivation. The phosphorylated forminteracts with PML. |
| 亚细胞定位 | Cytoplasm, cytoskeleton, spindle. Nucleus. Cytoplasm, cytoskeleton, centrosome. Cytoplasm. Note=Associated with the spindle duringprometaphase and metaphase. PEA15-binding andphosphorylated DAPK1 promote its cytoplasmic retention.Phosphorylation at Ser-244 and Ser-246 as well asautophosphorylation at Thr-188 promote nuclear localization. |
| 组织特异性 | Widely expressed. |
| 翻译后修饰 | Dually phosphorylated on Thr-183 and Tyr-185, which activatesthe enzyme. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-185. Phosphorylated upon FLT3 and KIT signaling. |
| 相似性 | Belongs to the protein kinase superfamily. CMGCSer/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. |
| 功能 | 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 by activated KIT and KITLG/SCF. Depending on the cellularcontext, the MAPK/ERK cascade mediates diverse biological functionssuch as cell growth, adhesion, survival and differentiation throughthe regulation of transcription, translation, cytoskeletalrearrangements. The MAPK/ERK cascade plays also a role ininitiation and regulation of meiosis, mitosis, and postmitoticfunctions in differentiated cells by phosphorylating a number oftranscription factors. About 160 substrates have already beendiscovered for ERKs. Many of these substrates are localized in thenucleus, and seem to participate in the regulation of transcriptionupon stimulation. However, other substrates are found in thecytosol as well as in other cellular organelles, and those areresponsible for processes such as translation, mitosis andapoptosis. Moreover, the MAPK/ERK cascade is also involved in theregulation of the endosomal dynamics, including lysosome processingand endosome cycling through the perinuclear recycling compartment(PNRC); as well as in the fragmentation of the Golgi apparatusduring mitosis. The substrates include transcription factors (suchas ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements(such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1),regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3,MCL1 or PPARG), regulators of translation (such as EIF4EBP1) and avariety of other signaling-related molecules (like ARHGEF2, DCC,FRS2 or GRB10). Protein kinases (such as RAF1, RPS6KA1/RSK1,RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1,MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) andphosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are othersubstrates which enable the propagation the MAPK/ERK signal toadditional cytosolic and nuclear targets, thereby extending thespecificity of the cascade. Mediates phosphorylation of TPR inrespons to EGF stimulation. May play a role in the spindle assemblycheckpoint. Phosphorylates PML and promotes its interaction withPIN1, leading to PML degradation (By similarity).
Acts as a transcriptional repressor. Binds to a[GC]AAA[GC] consensus sequence. Repress the expression ofinterferon gamma-induced genes. Seems to bind to the promoter ofCCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 andSTAT1. Transcriptional activity is independent of kinase activity. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | The protein encoded by this gene is a member of the MAPkinase family. MAP kinases, also known as extracellularsignal-regulated kinases (ERKs), act in a signaling cascade thatregulates various cellular processes such as proliferation,differentiation, and cell cycle progression in response to avariety of extracellular signals. This kinase is activated byupstream kinases, resulting in its translocation to the nucleuswhere it phosphorylates nuclear targets. Alternatively splicedtranscript variants encoding different protein isoforms have beendescribed. [provided by RefSeq, Jul 2008]. |
| 应用 | 推荐稀释比例 |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
| {Flow-Cyt} | {2ug/Test} |
Blank control (blue line): U251 (blue).
Primary Antibody (green line): Rabbit Anti-phospho-Erk1 (Thr202 + Tyr204) antibody (bs-1645R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 2% Paraformaldehyde (10 min)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Primary Antibody (green line): Rabbit Anti-phospho-Erk1 (Thr202 + Tyr204) antibody (bs-1645R)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 2% Paraformaldehyde (10 min)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-1645R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-1645R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Blank control:MCF7.
Primary Antibody (green line): Rabbit Anti-phospho-Erk1 (Thr202 + Tyr204) antibody (bs- 1645R)
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.
Primary Antibody (green line): Rabbit Anti-phospho-Erk1 (Thr202 + Tyr204) antibody (bs- 1645R)
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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相关实验
secondary antibody were purchased from GIBCO-BRL. Phosphospecific rabbit polyclonal antibodies against 180Thr and 182Tyr dual-phosphorylated p38, against total p38 (phosphorylation state independent), against 204Tyr-phosphorylated ERK1/2 and against total
(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
。所以，我们建议使用微波炉或高压锅加热煮沸以达到最佳抗原修复效果。抗体：Phospho–Stat3 (Tyr705)(D3A7) XP. Rabbit mAb #9145样本：石蜡包埋人类肺部肿瘤样本煮沸设备：浴锅 (左)，微波炉 (中)，高压锅 (右)免疫染色—封闭在 IHC–P 中，我们建议在室温下在含有 Tween20 的 TBST 缓冲液和 5% 正常山羊血清 (NGS) 中封闭样品，时长 1 小时，以防止非特异性背景染色。在进行 IHC–F 时，在含有 0.3% Triton. X–100 的 1X
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phospho-Erk1 (Thr202 + Tyr204) Rabbit pAb(bs-1645R)-50ul/100ul/200ul
￥1180 - 2800
