Invitrogen

Phospho-ERK1/ERK2 (Tyr204) Monoclonal Antibody (B.742.5)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View Details

3 Published Figures

1 Reference

View all (51) ERK1/2 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

Phospho-ERK1/ERK2 (Tyr204) Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Phospho-p44/42 MAPK/Erk1/2 pThr202/Tyr204 in NIH/3T3 cells stimulated with Platelet-Derived Growth Factor, using a Phospho-p44/42 MAPK/Erk1/2 pThr202/Tyr204 monoclonal antibody (Product # MA5-15174) (green). Actin filaments are labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.

Phospho-ERK1/ERK2 (Tyr204) Antibody in Western Blot (WB) — WB 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), NIH/3T3 (8) and NIH/3T3 treated with PMA (200 ng/mL for 10 minutes) (9). The blot was probed with Phospho-ERK1/ERK2 (Tyr204) Rabbit Monoclonal Antibody (Product # MA5-15174, 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 (Tyr204) was detected and increased upon EGF and PMA treatment across cell lines and pre-treatment with Afatinib (antagonist) resulted in inhibition of Phospho-ERK1/ERK2 (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 System (Product # EI0002) and Novex Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then wet transferred onto a nitrocellulose membrane. 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).

Phospho-ERK1/ERK2 (Tyr204) Antibody in Flow Cytometry (Flow) — Flow cytometric analysis of Phospho-p44/42 MAPK pErk1/2 pThr202/Tyr204 in U0126-inhibited (blue) or PMA-stimulated (green) Jurkat cells using a Phospho-p44/42 MAPK pErk1/2 pThr202/Tyr204 monoclonal antibody (Product # MA5-15174) compared to a nonspecific negative control antibody (red).

Phospho-ERK1/ERK2 (Tyr204) Antibody in Cell treatment — Altered expression of target protein upon TM demonstrates antibody specificity. Western blot analysis of Phospho-ERK1/ERK2 (Tyr204) using Anti-Phospho-ERK1/ERK2 (Tyr204) Monoclonal Antibody (Product # MA5-15174) shows increased expression of proteins phosphorylated at the tyrosine residues in A-431, MCF7,HeLa and NIH3T3 cell lines upon EGF, PMA and PDGF treatments. Pre-treatment with antagonist, Afatinib resulted in inhibition of ERK1/ERK2 (Tyr204) phosphorylation upon EGF treatment. {TM}

Phospho-ERK1/ERK2 (Tyr204) Antibody — Figure 2. Detection of ERK activity in corals. ( A ) Fluorescent immunoblot revealing activated (pERK) and total forms of ERK (ERK) present in Stylphora pistillata nubbins. Molecular weight standards in kilo Daltons (kDa) are indicated on the left side of the figure. ( B ) Immunoblot performed with ERK and pERK antibodies on protein extracts from coral nubbins incubated in the absence (Control) or presence of the MEK inhibitor U0126. Densitometric analysis of activated ERK intensities is presented on the right of the figure. The amido black total protein staining of the western blot membrane is shown as a loading control. The medians and standard deviations of three independent experiments are presented (***, p<0.01, t-test).

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1:200

Immunocytochemistry (ICC)

1:200

Immunofluorescence (IF)

1:200

Western Blot (WB)

1:1000

View 1 publication 1 publication

Product Specifications

Species Reactivity

Non-human primate, Pig, Rat, Zebrafish, Fruit fly, Human, Mink, Mouse

Published species

Not Applicable

Host / Isotype

Rabbit / IgG

Class

Monoclonal

Type

Antibody

Clone

B.742.5

Immunogen

Synthetic phosphopeptide corresponding to residues surrounding pThr202/Tyr204 of human p44 MAP kinase

Conjugate

Unconjugated

Form

Liquid

Purification

Affinity chromatography

Storage buffer

0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/mL BSA, 50% glycerol

Contains

<0.02% sodium azide

Storage conditions

-20°C

RRID

AB_10980347

Product Specific Information

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.

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.

Bioinformatics

Protein Aliases: CG12559-PA; CG12559-PC; CG12559-PD; CG12559-PE; CG12559-PG; CG12559-PH; CG12559-PI; Diphospho-ERK; drosophila ERK; enhancer of seven in absentia 7; ERK; Erk MAP kinase; ERK-1; ERK-2; ERK/rolled; ERK1; ERK2; ERT1; ERT2; extracellular signal-regulated kinase; Extracellular signal-regulated kinase 1; Extracellular signal-regulated kinase 2; extracellular signal-regulated kinase A; extracellular signal-regulated kinase-2; extracellular signal-related kinase 1; Extracellular-regulated kinase A; extracellular-signal-regulated kinase 2; Extracellular-signal-related kinase A; Insulin-stimulated MAP2 kinase; MAP kinase; MAP kinase 1; MAP kinase 2; MAP kinase 3; MAP kinase isoform p42; MAP kinase isoform p44; MAP-kinase; MAPK 1; MAPK 2; MAPK 3; MAPK1; MAPK3; Microtubule-associated protein 2 kinase; mitogen activated protein kinase; mitogen activated protein kinase 1; mitogen-activated 3; mitogen-activated protein kinase; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 2; Mitogen-activated protein kinase 3; Mitogen-activated protein kinase ERK-A; MNK1; p42-MAPK; p44 MAP kinase; p44-ERK1; p44-MAPK; phospho-ERK; pp42/MAP kinase; PRKM1; PRKM2; PRKM3; protein kinase; Protein rolled; protein tyrosine kinase ERK2; rl-PA; rl-PC; rl-PD; rl-PE; rl-PG; rl-PH; rl-PI; roll; rolled; sevenmaker

Gene Aliases: 12559; 9030612K14Rik; AA407128; AU018647; BcDNA:RE08694; C78273; CG12559; CG18732; CT34260; CT39192; D-ERK; DERK; DERK-A; dm-dpERK; Dmel\CG12559; Dmel_CG12559; DmErk; DmERK-A; DmERKA; DmMAPK; dp-ERK; DpERK; dpERK1; dpMAPK; Dsor2; E(sina)7; EC2-1; EK2-1; ERK; ERK-1; ERK-2; ERK-A; Erk/Map kinase; ERK1; ERK2; ERKA; ERT1; ERT2; Esrk1; EY2-2; fi06b09; GroupII; HS44KDAP; HUMKER1A; l(2)41Ac; l(2R)EMS45-39; MAP-k; MAPK; MAPK1; MAPK2; MAPK3; MNK1; Mtap2k; p38; p40; p41; p41mapk; p42-MAPK; P42MAPK; p44; p44-ERK1; p44-MAPK; P44ERK1; P44MAPK; pERK; pMAPK; PRKM1; PRKM2; PRKM3; RL; rl/ERK; rl/MAPK; rll; SEM; SR2-1; Su(Raf)2B; wu:fi06b09; zERK1; zERK2

UniProt ID: (Human) P28482, (Human) P27361, (Mouse) P63085, (Rat) P63086, (Rat) P21708, (Mouse) Q63844, (Fruit fly) P40417

Entrez Gene ID: (Pig) 100153927, (Human) 5594, (Human) 5595, (Mouse) 26413, (Rat) 116590, (Rat) 50689, (Mouse) 26417, (Zebrafish) 360144, (Zebrafish) 399480, (Fruit fly) 3354888