Analysis Note

ControlIncluded Positive Control Lysate: Catalog # 12-302, EGF-stimulated A431 lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg (20 µL) of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Research Sub CategoryMAP Kinases

Immunoprecipitation: Use 1-10 µg per immuno-precipitation.

Detect phospho-MAP Kinase1/2 using this Anti-phospho-MAP Kinase1/2 validated for use in WB, IP, ELISA & IC.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Mitogen-activated protein kinase 3 (UniProt: P27632; also known as EC:2.7.11.24, MAP kinase 3, MAPK 3. ERT2, Extracellular signal-regulated kinase 1, ERK-1, Insulin-stimulated MAP2 kinase, MAP kinase isoform p44, p44-MAPK, Microtubule-associated protein 2 kinase p44-ERK1) is encoded by the MAPK3 (also known as ERK1, PRKM3) gene (Gene ID: 5595) in human. ERK-1 is a serine/threonine kinase that acts as an essential component of the MAP kinase signal transduction pathway. It is expressed in different tissues and its highest levels have been observed in the nervous system. The MAPK/ERK cascade mediates several diverse biological processes such as cell growth, adhesion, survival, and differentiation through the regulation of transcription, translation, and cytoskeletal rearrangements. It also plays a role in initiation and regulation of meiosis, mitosis, and post-mitotic functions in differentiated cells by phosphorylating a few transcription factors. About 160 substrates have been described for ERKs. The protein kinase domain of ERK-1 is localized to amino acids 42-330 and the TXY motif is located in amino acids 202-204. ERK-1 is phosphorylated by MEK1 and MEK2 on threonine 202 and tyrosine 204 in response to external stimuli like insulin or nerve growth factor and both these phosphorylations are essential for its activity. Three isoforms of ERK-1 have been described that are produced by alternative splicing.

Immunogen

KLH-conjugated phosphopeptide containing the pTEpY motif of Erk1 and Erk2. Clone: 12D4

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified mouse monoclonal IgG1k lyophilized from 1 mL 2x PBS / 0.09 % Na-azide / PEG and Sucrose.
Reconstitute with 1 mL H2O (15 min, RT) for a final concentration of 100 µg/mL.

Thiophilic adsoption and size exclusion chromatography

Format: Purified

Quality

Evaluated by western blot on RIPA lysates from EGF-stimulated human A431 carcinoma cells.

Western Blot Analysis: 0.1-1.0 µg/mL of this antibody detected phospho-MAPK in RIPA lysates from EGFstimulated human A431 carcinoma cells.

Specificity

Specific for phosphorylated (activated) Erk1 and Erk2. Antibody recognition requires phosphorylation of both the threonine and tyrosine residues of Erk1/Erk2 and will not interact with the non-phosphorylated form of the proteins. Does not cross react with activated SAP Kinase 1 or 2.

Storage and Stability

Stable for 1 year at from date of receipt.

Target description

42-44 kDa