Invitrogen

Phospho-ERK1/ERK2 (Thr185, Tyr187) Polyclonal Antibody

Advanced Verification

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

22 Published Figures

51 References

View all (51) ERK1/2 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

Phospho-ERK1/ERK2 (Thr185, Tyr187) Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of ERK1/2 (pTpY185/187) showing staining in the cytoplasm and nucleus of paraffin-embedded human breast carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ERK1/2 (pTpY185/187) polyclonal antibody (Product # 44-680G) diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Phospho-ERK1/ERK2 (Thr185, Tyr187) Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of MDA-MB-231 with treatment of EGF(100 ng/mL for 15 mins) (Lane 1), MDA-MB-231 (Lane 2), U-87 MG (Lane 3),, SH-SY5Y (Lane 4) and HeLa (Lane 5). The blots were probed with Anti-Phospho-p44 MAPK + p42 MAPK pThr185 + pTyr187 Rabbit polyclonal Antibody (Product# 44680G, 1 in 1000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). Bands of 42 kDa and 44 kDa corresponding to Phospho-p44 MAPK + p42 MAPK pThr185 + pTyr187 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). 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 (Thr185, Tyr187) Antibody in Cell treatment — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot of Phospho-ERK1/ERK2 (Thr185, Tyr187) using Anti - ERK1/2 [pTpY185/187]Rabbit polyclonal Antibody (Product # 44-680G), shows increase in expression of Phospho-ERK1/ERK2 (Thr185, Tyr187upon treatment with EGF in MDA-MB-231 cells. {TM}

Phospho-ERK1/ERK2 (Thr185, Tyr187) Antibody — Published figure using Phospho-ERK1/ERK2 (Thr185, Tyr187) polyclonal antibody (Product # 44-680G)

Product Details

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:10-1:100

View 2 publications 2 publications

Western Blot (WB)

1:1000

View 33 publications 33 publications

Miscellaneous PubMed (Misc)

View 7 publications 7 publications

Immunocytochemistry (ICC)

View 3 publications 3 publications

Immunofluorescence (IF)

View 3 publications 3 publications

Immunohistochemistry (IHC)

View 2 publications 2 publications

Immunohistochemistry - Free Floating (IHC (Free))

View 1 publication 1 publication

Product Specifications

Species Reactivity

Bovine, C. elegans, Chicken, Fruit fly, Human, Mouse, Rat, Xenopus

Published species

C. elegans, Human, Mouse, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human ERK1 and 2 that contains threonine 202/185 and tyrosine 204/187. This region is conserved among many species including rat, mouse, cow, frog, snail, nematode, and fruit fly.

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20°C

RRID

AB_2533719

Product Specific Information

Purified from rabbit serum by sequential epitope-specific chromatography, this product contains enough material for 10 mini-blots. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the sites of phosphorylation to remove antibody that is reactive with non-phosphorylated ERK1 and 2. The final product is generated by affinity chromatography using an ERK1 and 2-derived peptide that is phosphorylated at threonine 202/185 and tyrosine 204/187, respectively, within the activation loop.

This antibody has been used in western blotting and previous lots have been used in immunostaining. Other applications may work but have not been tested. The positive control used in western analysis were PC12 cells +/- Sorbitol; NIH3T3 +/- PDGF; and HEK293 +/- UV.

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-related kinase 1; Extracellular-regulated kinase A; extracellular-signal-regulated kinase 2; Extracellular-signal-related kinase A; Insulin-stimulated MAP2 kinase; M phase MAP 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; MBP kinase; 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 1a; Mitogen-activated protein kinase 2; Mitogen-activated protein kinase 3; Mitogen-activated protein kinase ERK-A; MNK1; Myelin basic protein kinase; Myelin xP42 protein kinase; p42-MAPK; p44 MAP kinase; p44-ERK1; p44-MAPK; phospho-ERK; pp42/MAP kinase; PRKM1; PRKM2; PRKM3; 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; BOS_16809; BOS_22892; 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; GroupII; HS44KDAP; HUMKER1A; l(2)41Ac; l(2R)EMS45-39; MAP-k; MAPK; MAPK1; mapk1-a; mapk1-b; mapk1a; MAPK2; MAPK3; MNK1; mpk1; 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; XELAEV_18010534mg; xp42

UniProt ID: (Bovine) P46196, (Human) P28482, (Human) P27361, (Mouse) P63085, (Rat) P63086, (Rat) P21708, (Mouse) Q63844, (Xenopus) P26696, (Fruit fly) P40417

Entrez Gene ID: (Bovine) 327672, (Chicken) 373953, (Human) 5594, (Bovine) 531391, (Human) 5595, (Mouse) 26413, (Rat) 116590, (Rat) 50689, (Mouse) 26417, (Xenopus) 398985, (Fruit fly) 3354888