Invitrogen

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

8 Published Figures
13 References
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Protocols
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Datasheet
Protocols
Questions & Answers
Tech Support

Product Details

46-9109-42

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.06 µg)/test
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Product Specifications

Species Reactivity

Human, Mouse

Published species

Dog, 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

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 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

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 Best Protocols 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 (cat. 00-8222) (100 µL cell sample + 100 µL 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 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.

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Bioinformatics

Protein Aliases: ERK-1; ERK-2; erk1 erk2; ERK1b; 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 MAP Kinase; p42-MAPK; p44 MAP kinase; p44-ERK1; p44-MAPK; pp42/MAP kinase; protein tyrosine kinase ERK2

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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; 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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Molecular Function: non-receptor serine/threonine protein kinase

Function(s)
MAPK cascade activation of MAPK activity MAPK import into nucleus transcription, DNA-templated protein phosphorylation apoptotic process chemotaxis response to stress cellular response to DNA damage stimulus cell cycle signal transduction chemical synaptic transmission axon guidance positive regulation of cell proliferation fibroblast growth factor receptor signaling pathway response to toxic substance positive regulation of peptidyl-threonine phosphorylation regulation of phosphatidylinositol 3-kinase signaling viral process peptidyl-serine phosphorylation peptidyl-threonine phosphorylation sensory perception of pain cytosine metabolic process platelet activation positive regulation of cell migration thyroid gland development regulation of protein stability lipopolysaccharide-mediated signaling pathway positive regulation of telomere maintenance via telomerase regulation of stress-activated MAPK cascade mammary gland epithelial cell proliferation Fc-epsilon receptor signaling pathway Fc-gamma receptor signaling pathway involved in phagocytosis ERBB signaling pathway outer ear morphogenesis response to exogenous dsRNA response to estrogen negative regulation of cell differentiation positive regulation of translation positive regulation of transcription, DNA-templated thymus development T cell receptor signaling pathway B cell receptor signaling pathway regulation of sequence-specific DNA binding transcription factor activity regulation of cytoskeleton organization positive regulation of telomerase activity Bergmann glial cell differentiation long-term synaptic potentiation face development JAK-STAT cascade involved in growth hormone signaling pathway lung morphogenesis trachea formation labyrinthine layer blood vessel development cardiac neural crest cell development involved in heart development ERK1 and ERK2 cascade response to epidermal growth factor caveolin-mediated endocytosis regulation of Golgi inheritance cellular response to granulocyte macrophage colony-stimulating factor stimulus regulation of cellular response to heat positive regulation of telomere capping regulation of early endosome to late endosome transport positive regulation of protein phosphorylation transcription initiation from RNA polymerase I promoter protein complex assembly DNA damage induced protein phosphorylation phosphorylation arachidonic acid metabolic process BMP signaling pathway positive regulation of cyclase activity positive regulation of histone phosphorylation positive regulation of histone acetylation peptidyl-tyrosine autophosphorylation positive regulation of transcription from RNA polymerase II promoter cartilage development positive regulation of ERK1 and ERK2 cascade interleukin-1-mediated signaling pathway cellular response to mechanical stimulus positive regulation of xenophagy negative regulation of apolipoprotein binding organ morphogenesis response to lipopolysaccharide intracellular signal transduction cellular response to organic substance
Process(es)
phosphotyrosine binding DNA binding protein serine/threonine kinase activity MAP kinase activity protein binding ATP binding transcription factor binding RNA polymerase II carboxy-terminal domain kinase activity kinase activity phosphatase binding mitogen-activated protein kinase kinase kinase binding identical protein binding scaffold protein binding nucleotide binding protein kinase activity transferase activity transferase activity, transferring phosphorus-containing groups protein kinase binding
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