Phospho-ERK1/ERK2 (Thr202, Tyr204) Monoclonal Antibody (S.812.9)/Phospho-ERK1 / ERK2（Thr202，Tyr204）单克隆抗体（S.812.9）

货号：MA5-15173

规格：200 µL

价格：4880

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Synthetic phosphopeptide corresponding to residues

物种：其它

宿主：兔

抗体亚型：IgG

克隆号：S.812.9

荧光染料：其它

类型:	单抗	同型对照：
浓度：		用法：	1-3 µL(ChIP)；1:400(IHC (P))；1:100(IP)；1:1000(WB)

产品详细信息It is not recommended to aliquot this antibody.This antibody is not cross-reactive with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinase.靶标信息ERK1 and ERK2 are widely expressed and are involved in the regulation of meiosis, mitosis, and postmitotic functions in differentiated cells. Many different stimuli, including growth factors, cytokines, virus infection, ligands for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors and transforming agents, activate the ERK1 and ERK2 pathways. When growth factors bind to the receptor tyrosine kinase, Ras interacts with Raf, the serine/threonine protein kinase and activates it as well. Once actived, Raf phosphorylates serine residue in 2 further kinases, MEK1/2, which in turn phosphorylates tyrosine/threonine in extracellular-signal regulated kinase (ERK) 1/2. Upon activation, the ERKs either phosphorylate a number of cytoplasmic targets or migrate to the nucleus, where they phosphorylate and activate a number of transcription factors such as c-Fos and Elk-1.

数据：

Phospho-ERK1/ERK2 (Thr202, Tyr204) Antibody (MA5-15173) in IHCImmunohistochemistry was performed on paraffin-embedded human colon carcinoma tissue. To expose target proteins, antigen retrieval was performed by boiling tissue sections in 10mM sodium citrate buffer, pH 6.0 for 10 minutes. Following antigen retrieval, endogenous peroxidase activity was quenched with 3% hydrogen peroxide for 10 minutes, tissues were blocked in 5% normal goat serum in TBST for 1 hour at room temperature, and then probed with a Phospho-p44/42 MAPK (Erk1/2) (pThr202/Tyr204) monoclonal antibody (Product # MA5-15173) at a dilution of 1:400 overnight at 4°C. Detection was performed using an HRP-conjugated anti-rabbit IgG secondary reagent system followed by DAB substrate. Tissues were counterstained with Hematoxylin and visualized by light microscopy. Phospho-ERK1/ERK2 (Thr202, Tyr204) Antibody (MA5-15173) in WBWB analysis was performed on whole cell extracts (30 µg) of A-431 (1), A-431 treated with EGF (200 ng/mL for 10 minutes) (2), A-431 treated with Afatinib followed by EGF (0.5uM of Afatinib for 6hrs, 200 ng/mL for 10 minutes) (3), MCF7 (4) and MCF7 treated with PMA (200 ng/mL for 10 minutes) (5), HeLa (6) and HeLa treated with PMA (200 ng/mL for 10 minutes) (7). The blot was probed with Phospho-ERK1/ERK2 (Thr202, Tyr204) Rabbit Monoclonal Antibody (Product # MA5-15173, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 1:4000 dilution). 44, 42 kDa band corresponding to Phospho-ERK1/ERK2 (Thr202, Tyr204) was detected and increased upon EGF and PMA treatment across cell lines tested and pre-treatment with Afatinib (antagonist) resulted in inhibition of Phospho-ERK1/ERK2 (Thr202, Tyr204) in A-431 cell line upon EGF treatment. Protein samples were electrophoresed using Novex NuPAGE 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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参考文献：

1. BMC cancerRas-ERK1/2 signaling contributes to the development of colorectal cancer via regulating H3K9ac.

技术参数

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