Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2) antibody produced in mouse
SIGMA/M8159 - clone MAPK-YT, ascites fluid
Synonym: Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2)
Product Type: Chemical
Western Blotting JOURNAL CITATION: Claudin-3 overexpression increases the malignant potential of colorectal cancer cells: roles of ERK1/2 and PI3K-Akt as modulators of EGFR signaling. By: de Souza, W. F., Fortunato-Miranda, N., et al. in PLoS One, 2013. PubMed ID: 24069372 Image collected and cropped by CiteAb from the following publication, (http://dx.plos.org/10.1371/journal.pone.0074994), provided under a CC-BY license. Image might reflect a new usage that is not reflected in claims on product description or independently verified by Merck KGaA, Darmstadt, Germany. For Research Use Only. Not for use in diagnostic procedures.
Immunofluorescence
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase, Activated (Cat No. M8159) (green). The activated state of MAP kinase signaling pathways trigggered by receptor tyrosine kinases can be followed in situ, revealing the time and amount of activation, the extent of ligand diffusion, and the effects of inhibitory ligands.
From L. Gabay, R. Seger, B-Z. Shilo, Weizmann Institute of Science, Rehovot, Israel; reproduced cover photograph from Science, 277, 1103 (1997). Used with permission.
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase, Activated (Cat No. M8159) (green). The activated state of MAP kinase signaling pathways trigggered by receptor tyrosine kinases can be followed in situ, revealing the time and amount of activation, the extent of ligand diffusion, and the effects of inhibitory ligands.
From L. Gabay, R. Seger, B-Z. Shilo, Weizmann Institute of Science, Rehovot, Israel; reproduced cover photograph from Science, 277, 1103 (1997). Used with permission.
Monoclonal Anti-MAP Kinase, Activated, Clone MAPK-YT (M 8159)
Immunoblot
Extract of Rat1 cells activated with 100μM Vanadate+200μM H2O2 was separated on SDS-PAGE and blotted with Monoclonal Anti-MAP Kinase, Activated (Product No. M 8159). The antibody was developed with Goat Anti-Mouse IgG, AlkPhos Conjugate (Product No. A 1293) and NBT/BCIP substrate.
Lane A: Antibody dilution 1:10,000
Lane B: Negative control (only secondary antibody)
Immunoblot
Extract of Rat1 cells activated with 100μM Vanadate+200μM H2O2 was separated on SDS-PAGE and blotted with Monoclonal Anti-MAP Kinase, Activated (Product No. M 8159). The antibody was developed with Goat Anti-Mouse IgG, AlkPhos Conjugate (Product No. A 1293) and NBT/BCIP substrate.
Lane A: Antibody dilution 1:10,000
Lane B: Negative control (only secondary antibody)
Immunofluorescence NRK cells were fixed and permeabilized with methanol followed by methanol:acetone. Fixed cells were stained with 1:1,000 Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2) Clone: MAPK-YT (Cat. No. M8159). The antibody was developed using Goat Anti-Mouse IgG, Cy3™ conjugate. Cells were counterstained with DAPI (blue) to stain nuclei.
| antibody form | ascites fluid |
| antibody product type | primary antibodies |
| biological source | mouse |
| clone | MAPK-YT, monoclonal |
| conjugate | unconjugated |
| contains | 15 mM sodium azide |
| isotype | IgG1 |
| mol wt | antigen ERK-1 44 kDa |
| antigen ERK-2 42 kDa | |
| Quality Level | 200  |
| shipped in | dry ice |
| species reactivity | human, Caenorhabditis elegans, Xenopus, Drosophila, hamster, rat, bovine, mouse, yeast |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunocytochemistry: suitable |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| immunoprecipitation (IP): suitable | |
| indirect ELISA: suitable | |
| western blot: 1:10,000 using rat brain extract | |
| UniProt accession no. | P28482  |
| Application: | Antibodies that react specifically with the active form of MAP kinase are useful for the study of the specific activation requirements, differential tissue expression, and intracellular localization of the active form of MAP kinase in normal and neoplastic tissue. Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2) may be used for the localization of the active, dually-phosphorylated, form of MAP kinase using various immunochemical assays such as immunoblotting of cultured cells and tissue extracts, ELISA, immunocytochemistry, immunoprecipitation, and in immunohistochemistry (formalin and formaldehyde-fixed sections). Reactivity has been observed with human, bovine, rat, mouse, Drosophila, Spodoptera frugiperda, and yeast. Monoclonal Anti-MAP Kinase, activated (Diphosphorylated ERK-1&2) antibody has been used in plasmid and transient transfection and western blotting. |
| Biochem/physiol Actions: | MAP kinase (MAPK, mitogen-activated protein kinase) plays a crucial role in various signal transduction pathways leading signals of growth factor, as well as G protein-coupled receptors to their intracellular targets. MAP kinase was shown to regulate several cellular processes among them proliferation, differentiation, cellular morphology and oncogenesis. Activation of ERK-1 and ERK-2 in mitogen-stimulated cells is directly mediated by MAP kinase kinase (MAPKK or MEK), a dual-specificity protein kinase, which phosphorylates both threonine and tyrosine residues in the regulatory sites of MAP kinase. Following activation, MAP kinase phosphorylates several nuclear targets, including transcription factors as well as membrane and cytoskeletal proteins. Termination of MAP kinase signalling appears to be mediated by MAP kinase phosphatase, MKP-1, a dual specificity Thr/Tyr phosphatase which dephosphorylates and inactivates MAP kinase. |
| Biochem/physiol Actions: | The antibody reacts specifically with the diphosphorylated form of MAP kinase (ERK-1 and ERK-2). It does not recognize the non-phosphorylated or the monophosphorylated forms of MAP kinase or the diphosphorylated forms of JNK and p38 MAP kinase. The epitope recognized by the antibody contains the phosphorylated threonine and tyrosine residues within the regulatory site of active MAP kinase. |
| 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: | MAP kinase (MAPK, mitogen-activated protein kinase is also termed as ERK, extracellular regulated protein kinase). Molecular cloning has established that MAP kinase (ERKs) consists of at least four isoforms: ERK-1 (p44mapk), ERK-2 (p42mapk), ERK-3, and ERK-5. MAP kinase isoforms appear to be widely expressed in the central nervous system, thymus, spleen, heart, lung and kidney. It is also expressed at high levels in PC12 cells and in fibroblasts. MAPK1 is located on human chromosome 22q11. MAPK3 is mapped to human chromosome 16p11. |
| Immunogen: | synthetic peptide HTGFLpTEpYVAT corresponding to the phosphorylated form of the ERK-activation loop. |
| Physical form: | The product is provided as ascites fluid containing 15 mM sodium azide as a preservative. |
| Preparation Note: | Store at 2-8 °C for up to one month. For extended storage, the solution may be frozen in working aliquots. Repeated freezing and thawing is not recommended. Storage in "frost-free" freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |