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Invitrogen

Phospho-ERK1/2 (Thr202, Tyr204) Monoclonal Antibody (MILAN8R), PerCP-eFluor™ 710, eBioscience™

8 Published Figures
14 References
View all (75) ERK1/2 antibodies

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Product Details

46-9109-42

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.06 µg)/test
View 14 publications 14 publications
Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

MILAN8R

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2573872

Product Specific Information

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 and flow cytometric analysis of stimulated normal human peripheral blood cells or stimulated mouse splenocytes. This can be used at 5 µL (0.06 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.

Protocols: 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 "Staining intracellular Antigens for Flow Cytometry Protocol" located in the BestProtocols® Section under the Resources tab online.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

Bioinformatics

Protein Aliases: ERK-1; ERK-2; erk1 erk2; Erk1; cancer susceptibility protein; ERK1b; ERK1c; Erk2; cancer susceptibility protein; ERT1; ERT2; Extracellular signal-regulated kinase 1; Extracellular signal-regulated kinase 2; extracellular signal-related kinase 1; Insulin-stimulated MAP2 kinase; MAP kinase 1; MAP kinase 2; MAP kinase 3; MAP kinase isoform p42; MAP kinase isoform p44; MAPK 1; MAPK 2; Microtubule-associated protein 2 kinase; mitogen activated protein kinase 1; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 2; Mitogen-activated protein kinase 3; MNK1; p42; p42 MAP Kinase; p42-MAPK; p44 MAP kinase; p44-ERK1; p44-MAPK; pp42/MAP kinase; protein tyrosine kinase ERK2; unnamed protein product

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Gene Aliases: 9030612K14Rik; AA407128; AU018647; C78273; ERK; ERK-1; ERK-2; ERK1; ERK2; ERT1; ERT2; Esrk1; HS44KDAP; HUMKER1A; Mapk; MAPK1; MAPK2; MAPK3; Mnk1; Mtap2k; NS13; p38; p40; p41; p41mapk; p42-MAPK; P42MAPK; p44; p44-ERK1; p44-MAPK; P44ERK1; P44MAPK; PRKM1; PRKM2; PRKM3

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UniProt ID: (Human) P28482, (Human) P27361, (Mouse) P63085, (Mouse) Q63844

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Entrez Gene ID: (Human) 5594, (Human) 5595, (Mouse) 26413, (Mouse) 26417

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Function(s)
nucleotide binding phosphotyrosine binding DNA binding protein kinase activity protein serine/threonine kinase activity MAP kinase activity protein binding ATP binding RNA polymerase II carboxy-terminal domain kinase activity kinase activity transferase activity phosphatase binding identical protein binding protein serine kinase activity DNA-binding transcription factor binding transcription factor binding transferase activity, transferring phosphorus-containing groups protein kinase binding mitogen-activated protein kinase kinase kinase binding scaffold protein binding non-receptor serine/threonine protein kinase
Process(es)
MAPK cascade regulation of transcription from RNA polymerase II promoter protein phosphorylation apoptotic process chemotaxis cellular response to DNA damage stimulus signal transduction cell surface receptor signaling pathway epidermal growth factor receptor signaling pathway chemical synaptic transmission heart development learning or memory insulin receptor signaling pathway organ morphogenesis positive regulation of macrophage chemotaxis positive regulation of peptidyl-threonine phosphorylation neural crest cell development Schwann cell development peptidyl-threonine phosphorylation cytosine metabolic process regulation of ossification regulation of cellular pH thyroid gland development regulation of protein stability lipopolysaccharide-mediated signaling pathway positive regulation of telomere maintenance response to lipopolysaccharide regulation of stress-activated MAPK cascade mammary gland epithelial cell proliferation cellular response to amino acid starvation response to nicotine intracellular signal transduction ERBB signaling pathway ERBB2-ERBB3 signaling pathway outer ear morphogenesis myelination response to exogenous dsRNA positive regulation of cholesterol biosynthetic process negative regulation of cell differentiation insulin-like growth factor receptor signaling pathway thymus development T cell receptor signaling pathway B cell receptor signaling pathway stress-activated MAPK cascade regulation of cytoskeleton organization Bergmann glial cell differentiation long-term synaptic potentiation face development lung morphogenesis trachea formation labyrinthine layer blood vessel development cardiac neural crest cell development involved in heart development interleukin-34-mediated signaling pathway chemokine-mediated signaling pathway ERK1 and ERK2 cascade response to epidermal growth factor cellular response to tumor necrosis factor caveolin-mediated endocytosis regulation of Golgi inheritance positive regulation of macrophage proliferation positive regulation of neuroinflammatory response regulation of early endosome to late endosome transport autophagosome assembly negative regulation of T cell mediated immune response to tumor cell transcription, DNA-templated regulation of gene expression negative regulation of autophagy positive regulation of autophagy phosphorylation sensory perception of pain BMP signaling pathway positive regulation of cyclase activity cellular response to nutrient levels negative regulation of proteasomal ubiquitin-dependent protein catabolic process peptidyl-tyrosine autophosphorylation TORC1 signaling glucose homeostasis signal transduction in response to DNA damage positive regulation of transcription from RNA polymerase II promoter modulation of synaptic transmission protein stabilization negative regulation of T cell activation cartilage development positive regulation of ERK1 and ERK2 cascade interleukin-1-mediated signaling pathway cellular response to mechanical stimulus negative regulation of cholesterol efflux xenophagy negative regulation of TORC1 signaling positive regulation of xenophagy MAPK import into nucleus cell cycle response to toxic substance peptidyl-serine phosphorylation response to estrogen positive regulation of translation regulation of sequence-specific DNA binding transcription factor activity cellular response to organic substance cellular response to granulocyte macrophage colony-stimulating factor stimulus positive regulation of protein phosphorylation protein complex assembly DNA damage induced protein phosphorylation positive regulation of histone phosphorylation positive regulation of histone acetylation negative regulation of apolipoprotein binding
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