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Rabbit Anti-phospho-ERK1/2 (Thr202 + Tyr204)  antibody (bs-3016R)
~~~促销，代码KX240301~~~
~~~促销，代码KX240302~~~
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产品编号 bs-3016R
英文名称 phospho-ERK1/2 (Thr202 + Tyr204)
中文名称 磷酸化丝裂原活化蛋白激酶1/2抗体
别    名 ERK1 (phospho T202 + Y204); p-ERK1 (phospho T202 + Y204); Erk1 (pT202/pY204) + Erk2 (pT185/pY187); p-ERK1/2(T202/Y204); ERK 1; ERK 2; MK03_HUMAN; MK01_HUMAN; ERK-2; ERK1; ERK2; ERT1; ERT2; Extracellular signal regulated kinase 1; Extracellular signal regulated kinase 1; Extracellular signal regulated kinase 2; Extracellular signal regulated kinase 2; Extracellular signal-regulated kinase 2; HS44KDAP; HUMKER1A; Insulin stimulated MAP2 kinase; MAP kinase 1; MAP kinase 2; MAP kinase isoform p42; MAP kinase isoform p44; MAPK 1; MAPK 2; MAPK1; MAPK2; MGC20180; Microtubule associated protein 2 kinase; Mitogen activated protein kinase 1; Mitogen activated protein kinase 1; Mitogen activated protein kinase 2; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 2; MK01_MOUSE; p38; p40; p41; p41mapk; p42 MAPK; p42-MAPK; p42MAPK; p42MAPK; p44 ERK1; p44 MAPK; p44ERK1; p44ERK1; p44MAPK; p44MAPK; PRKM 1; PRKM 1; PRKM 2; PRKM 2; PRKM1; PRKM2; Protein kinase mitogen activated 1; Protein kinase mitogen activated 1; Protein kinase mitogen activated 2; Protein kinase mitogen activated 2; Protein tyrosine kinas.  
Specific References  (68)     |     bs-3016R has been referenced in 68 publications.
[IF=14.528] Dejun Xu. et al. Melatonin protects mouse testes from palmitic acid‐induced lipotoxicity by attenuating oxidative stress and DNA damage in a SIRT1‐dependent manner. J Pineal Res. 2020 Nov;69(4):e12690  WB ;  Mouse.  
[IF=10.633] Fanglin Wang. et al. Adipose-derived stem cells with miR-150-5p inhibition laden in hydroxyapatite/tricalcium phosphate ceramic powders promote osteogenesis via regulating Notch3 and activating FAK/ERK and RhoA. ACTA BIOMATER. 2022 Oct;:  WB, IF ;  Human.  
[IF=9.078] Tzu-Hsiang Lin. et al. A bilineage thermosensitive hydrogel system for stimulation of mesenchymal stem cell differentiation and enhancement of osteochondral regeneration. Compos Part B-Eng. 2022 Jan;:109614  ICC ;  Rabbit.  
[IF=8.109] Li X et al. Disseminated Melanoma Cells Transdifferentiate into Endothelial Cells in Intravascular Niches at Metastatic Sites. Cell Rep. 2020 Jun 16;31(11):107765.  IHF ;  Mouse.  
[IF=8.039] Yifan Zhu. et al. Discovery of Selective P2Y6R Antagonists with High Affinity and In Vivo Efficacy for Inflammatory Disease Therapy. J MED CHEM. 2023;XXXX(XXX):XXX-XXX  WB ;  Mouse.  
[IF=7.675] Honghong Zhan. et al. Oxybaphus himalaicus Mitigates Lipopolysaccharide-Induced Acute Kidney Injury by Inhibiting TLR4/MD2 Complex Formation. ANTIOXIDANTS-BASEL. 2022 Dec;11(12):2307  WB ;  Mouse.  
[IF=7.129] Jin Chen. et al. Surface functionalization-dependent inflammatory potential of polystyrene nanoplastics through the activation of MAPK/ NF-κB signaling pathways in macrophage Raw 264.7. ECOTOX ENVIRON SAFE. 2023 Feb;251:114520  WB ;  Mouse.  
[IF=7.076] Yajing Lv. et al. Nucleotide de novo synthesis increases breast cancer stemness and metastasis via cGMP-PKG-MAPK signaling pathway. Plos Biol. 2020 Nov;18(11):e3000872  WB ;  Mouse.  
[IF=6.317] Shiqing Sun. et al. Pharmacodynamic structure of deer antler base protein and its mammary gland hyperplasia inhibition mechanism by mediating Raf-1/MEK/ERK signaling pathway activation. FOOD FUNCT. 2023 Mar;:  WB ;  Rat.  
[IF=6.126] Xiaosu Chen. et al. DCBLD2 Mediates Epithelial-Mesenchymal Transition-Induced Metastasis by Cisplatin in Lung Adenocarcinoma. Cancers. 2021 Jan;13(6):1403  WB ;  Human.  
[IF=5.455] Zhang, Rongrong. et al. Compound traditional Chinese medicine dermatitis ointment ameliorates inflammatory responses and dysregulation of itch-related molecules in atopic dermatitis. Chin Med-Uk. 2022 Dec;17(1):1-19  WB ;  Mouse.  
[IF=5.31] Ning Zhong. et al. Apatinib inhibits the growth of small cell lung cancer via a mechanism mediated by VEGF, PI3K/Akt and Ki-67/CD31. 2021 Sep 30  IF ;  mouse.  
[IF=5.279] Lei Wang. et al. Lactobacillus plantarum DP189 Reduces α-SYN Aggravation in MPTP-Induced Parkinson’s Disease Mice via Regulating Oxidative Damage, Inflammation, and Gut Microbiota Disorder. J Agr Food Chem. 2022;70(4):1163–1173  WB ;  Mouse.  
[IF=5.279] Wei Liu. et al. Glycolysis and Reactive Oxygen Species Production Participate in T-2 Toxin-Stimulated Chicken Heterophil Extracellular Traps. J Agr Food Chem. 2021;XXXX(XXX):XXX-XXX  WB ;  Chicken.  
[IF=5.168] Guo-Jian Jiang. et al. Carteolol triggers senescence via activation of β-arrestin–ERK–NOX4–ROS pathway in human corneal endothelial cells in vitro. CHEM-BIOL INTERACT. 2023 Apr;:110511  WB ;  Human.  
[IF=5.153] Jiang, Liqiang. et al. Chicken heterophils extracellular traps act as early effectors against cyclopiazonic acid dependent upon NADPH oxidase, ROS and glycolysis. ARCH TOXICOL. 2022 May;:1-10  WB ;  Chicken.  
[IF=5.008] Tan et al. ART3 regulates triple-negative breast cancer cell function via activation of Akt and ERK pathways. (2016) Oncotarge. 7:46589-46602  WB,IHC ;  Human.  
[IF=4.996] Cong Changsheng. et al. Renin-angiotensin system inhibitors mitigate radiation pneumonitis by activating ACE2-angiotensin-(1–7) axis via NF-κB/MAPK pathway. SCI REP-UK. 2023 May;13(1):1-11  WB ;  Mouse.  
[IF=4.75] Rosenzweig, Derek H., Sing J. Ou, and Thomas M. Quinn. ʺP38 mitogen activated protein kinase promotes dedifferentiation of primary articular chondrocytes in monolayer culture.ʺ Journal of Cellular and Molecular Medicine (2013).  WB ;  Bovine.  
[IF=4.658] Zhang J et al. Down‐regulation of Suv39h1 attenuates neointima formation after carotid artery injury in diabetic rats. J Cell Mol Med. 2019 Nov 17.  WB ;  Rat.  
[IF=4.486] Jianye Yang. et al. Astragalus polysaccharide attenuates LPS-related inflammatory osteolysis by suppressing osteoclastogenesis by reducing the MAPK signalling pathway. 2021 Jun 02  WB ;  Mouse.  
[IF=4.411] Yudan Zhao. et al. COX-2 is required to mediate crosstalk of ROS-dependent activation of MAPK/NF-κB signaling with pro-inflammatory response and defense-related NO enhancement during challenge of macrophage-like cell line with Giardia duodenalis. PLOS NEGLECT TROP D. 2022 Apr;16(4):e0010402  WB ;  Mouse.  
[IF=4.225] Qihe Tang. et al. Bergenin Monohydrate Attenuates Inflammatory Response via MAPK and NF-κB Pathways Against Klebsiella pneumonia Infection. Front Pharmacol. 2021; 12: 651664  WB ;  Mouse.  
[IF=4.2] Zhang, Fengwei, et al. "Seasonal Expression of Oxytocin and Oxytocin Receptor in the Scented Gland of Male Muskrat (Ondatra zibethicus)." Scientific Reports 7.1 (2017): 16627.  IHC-P ;  Others.  
[IF=4.171] Gu F et al. Lactobacillus casei improves depression-like behavior in chronic unpredictable mild stress-induced rats by BDNF-TrkB signal pathway and the intestinal microbiota. Food Funct . 2020 Jul 1;11(7):6148-6157.  WB ;  Rat.  
[IF=4.171] Gu F et al. Lactobacillus casei improves depression-like behavior in chronic unpredictable mild stress-induced rats by the BDNF-TrkB signal pathway and the intestinal microbiota. Food Funct. 2020 Jul 1;11(7):6148-6157.  WB ;  Rat.  
[IF=4.122] Jeon et al. Activation of the complement system in an osteosarcoma cell line promotes angiogenesis through enhanced production of growth factors. (2018) Sci.Rep. 8:5415  WB ;  
[IF=4.105] Ai Jin. et al. TSLP-induced collagen type-I synthesis through STAT3 and PRMT1 is sensitive to calcitriol in human lung fibroblasts. Bba-Mol Cell Res. 2021 Jun;:119083  WB ;  Human.  
[IF=3.943] Xiuhong Wang. et al. Protective effect of combination of anakinra and MCC950 against acute lung injury is achieved through suppression of the NF-κB-mediated-MAPK and NLRP3-caspase pathways. Int Immunopharmacol. 2021 Aug;97:107506  WB ;  Mouse.  
[IF=3.887] Yan LI. et al. The combination of EGCG with warfarin reduces deep vein thrombosis in rabbits through modulating HIF-1α and VEGF via the PI3K/AKT and ERK1/2 signaling pathways. CHIN J NAT MEDICINES. 2022 Sep;20:679  WB ;  Human.  
产品类型 磷酸化抗体 
研究领域 免疫学  神经生物学  信号转导  干细胞  激酶和磷酸酶  
抗体来源 Rabbit
克隆类型 Polyclonal
交叉反应 Rat,Mouse,Human (predicted: GuineaPig,Rabbit,Horse,Cow,Pig,Dog,Chicken)
产品应用 WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC=1:100, IF=1:100-500, Flow-Cyt=1μg /test, ELISA=1:5000-10000
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
理论分子量 41kDa
细胞定位 细胞核 细胞浆 
性    状 Liquid
浓    度 1mg/ml
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from mouse p44/42 MAPK around the phosphorylation site of Thr202/Tyr204: FL(p-T)E(p-Y)VA 
亚    型 IgG
纯化方法 affinity purified by Protein A
缓 冲 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存条件 Shipped at 4℃. Store at -20 °C 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.
PubMed PubMed
产品介绍 p44/42 MAP Kinase(Phospho-Thr202); ERK (extracellular signal regulated kinase), also known as MAPK (mitogen activated protein kinase) has two closely related isoforms of 44 kDa and 42 kDa, respectively. These kinases belong to a family of serine/threonine kinases that are activated upon treatment of cells with a large variety of stimuli including mitogens, hormones, growth factors, cytokines, and bioactive peptides. Cell stimulation induces the activation of a signaling cascade, the downstream effects of which have been linked to the regulation of cell growth and differentiation as well as the cytoskeleton. ERK1 and ERK2 are phosphorylated within the activation loop on both a Threonine and a Tyrosine residue (within a Thr-Glu-Tyr motif) by MEKs (MAPK/ERK kinases), thereby greatly elevating the activity of ERK1&2.

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 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). [FUNCTION] 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(By similarity).

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(via C-terminus region and phosphorylated form); the interactionrequires dimerization of MAPK1/ERK2 and increases following EGFstimulation (By similarity). Interacts (phosphorylated form) withCAV2 ('Tyr-19'-phosphorylated form); the interaction, promoted byinsulin, leads to nuclear location and MAPK1 activation (Bysimilarity). Interacts with DCC (By similarity). Interacts withMORG1, PEA15 and MKNK2. MKNK2 isoform 1 binding prevents fromdephosphorylation and inactivation. The phosphorylated forminteracts with PML (By similarity).

Subcellular Location:
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 (Bysimilarity).

Tissue Specificity:
Widely expressed.

Post-translational modifications:
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.

Similarity:
Belongs to the protein kinase superfamily. CMGCSer/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain.

SWISS:
P27361

Gene ID:
5595

Database links:

Entrez Gene: 5594 Human

Entrez Gene: 5595 Human

Entrez Gene: 26413 Mouse

Entrez Gene: 26417 Mouse

Entrez Gene: 116590 Rat

Entrez Gene: 50689 Rat



激酶和磷酸酶（Kinases and Phosphatases）
丝裂原活化蛋白激酶-ERK1/2（Mitogen-activated protein kinase 1/2)是一组可以被多种细胞外信号即获得蛋白丝/苏氨酸激酶，处于胞浆信号传导通路的终末位置，活化后转位到核内，作用于核内转录因子，调节基因表达。它主要参与生长因子、激素、细胞因子、应激等各种刺激下细胞的反应、细胞的生长、分化过程。
产品图片
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human Glioblastoma); 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:2000 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 2 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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 2 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R ) 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 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R ) 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 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-ERK12 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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+2 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:400 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+2 (Thr202 + Tyr204)) Polyclonal Antibody, Unconjugated (bs-3016R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-ERK1/2 (Thr202 + Tyr204)) polyclonal Antibody, Unconjugated (bs-3016R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Tissue/cell: Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-ERK1/2 (Thr202 + Tyr204)) polyclonal Antibody, Unconjugated (bs-3016R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Tissue/cell: human lung carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-phospho-ERK1/2(Thr202+Tyr204) Polyclonal Antibody, Unconjugated(bs-3016R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
Blank control (blue line): U251 (fixed with 2% paraformaldehyde (10 min)and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-phospho-ERK12 (Thr202 + Tyr204) antibody (bs-3016R),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.
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