Phospho-ERK1/2 (Thr202, Tyr204) Monoclonal Antibody (MILAN8R), PE-eFluor 610

货号：61-9109-42

规格：100 Tests

价格：3513

产品类型：流式抗体

品牌：Thermo Fisher

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG1, kappa

克隆号：MILAN8R

荧光染料：PE-eFluor 610

类型:	单抗	同型对照：	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PE-eFluor 610
浓度：	5 µL/Test	用法：	5 µL (0.5 µg)/test(Flow)

产品详细信息Description: This MILAN8R monoclonal antibody recognizes human and mouse extracellular signal-regulated kinases 1 and 2 (also known as ERK1/2, p44/p42, or MAPK3/1) when phosphorylated on T202/Y204. ERK1/2 belong to a family of conserved serine/threonine protein kinases known as mitogen-activated protein kinases (MAPKs) that are involved in many cellular programs such as proliferation, differentiation, motility, and survival. ERK1/2 signaling is activated in response to numerous extracellular stimuli including mitogens, growth factors, and cytokines. The primary activators of ERK1/2 are MEK1 and MEK2 which act by phosphorylating the activation loop residues T202/Y204 and T185/Y187 in ERK1 and ERK2, respectively. Several downstream targets of ERK1/2 have been identified, including p90RSK and the transcription factor Elk-1. ERK1/2 are negatively regulated by MAPK phosphatases, known as DUSPs or MKPs, as well as by chemical inhibitors of MEK including U0126 and PD98059. Disruption of the ERK pathway is common in many types of cancer.Specificity of this MILAN8R clone was determined by ELISA, flow cytometry, and western blotting.Applications Reported: This MILAN8R antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This MILAN8R antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.Staining Protocol: We recommend using Protocol C: Two-step protocol: Fixation/Methanol. Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins and Protocol B: One-step protocol: intracellular (nuclear) proteins cannot be used. All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2 µm post-manufacturing filtered.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-ERK1/2 (Thr202, Tyr204) Antibody (61-9109-42) in FlowTOP: Mouse splenocytes were unstimulated (left) or stimulated with F (ab')2 Anti-Mouse IgM, u chain specific Functional Grade Purified (Product # 16-5092-85) and Anti-Mouse CD40 Functional Grade Purified (Product # 16-0401-82) (right). The cells were then intracellularly stained with Anti-Human/Mouse CD45R (B220) FITC (Product # 11-0452-82) and Anti-Human/Mouse phospho-ERK1/2 (T202/Y204) PE-eFluor® 610 using the IC Fixation/Methanol Protocol. Cells in the lymphocyte gate were used for analysis. BOTTOM: Mouse splenocytes were unstimulated (orange histogram) or stimulated with F (ab')2 Anti-Mouse IgM, u chain specific Functional Grade Purified (Product # 16-5092-85) and Anti-Mouse CD40 Functional Grade Purified (Product # 16-0401-82) (purple histogram). The cells were then intracellularly stained with Anti-Human/Mouse CD45R (B220) FITC (Product # 11-0452-82) and Anti-Human/Mouse phospho-ERK1/2 (T202/Y204) PE-eFluor® 610 using the IC Fixation/Methanol Protocol. B220+ cells in the lymphocyte gate were used for analysis.

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参考文献：

1.Acta neuropathologicaGermline and somatic FGFR1 abnormalities in dysembryoplastic neuroepithelial tumors."Published figure using Phospho-ERK1/2 (Thr202, Tyr204) monoclonal antibody (Product # 61-9109-42) in Flow Cytometry"2.The Journal of clinical investigationHeterogeneity of leukemia-initiating capacity of chronic myelogenous leukemia stem cells."Published figure using Phospho-ERK1/2 (Thr202, Tyr204) monoclonal antibody (Product # 61-9109-42) in Flow Cytometry"3.Journal of immunology (Baltimore, Md. : 1950)The Bacterial Enzyme IdeS Cleaves the IgG-Type of B Cell Receptor (BCR), Abolishes BCR-Mediated Cell Signaling, and Inhibits Memory B Cell Activation."Published figure using Phospho-ERK1/2 (Thr202, Tyr204) monoclonal antibody (Product # 61-9109-42) in Flow Cytometry"

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