Immunofluorescence analysis of ERK1 was done on 70% confluent log phase U2OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ERK1 Mouse Monoclonal Antibody (138600) at 2 µg - 4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Panel d is a merged image showing nuclear and cytoplasmic localization of ERK1. Panel e shows no primary antibody. The images were captured at 20X magnification.
|Tested species reactivity||Dog, Human, Mouse, Rat|
|Published species reactivity||Tag, Pig, Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic Peptide corresponding to a section of the C-terminus of rat ERK1.|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
ERK2 mediates inner hair cell survival and decreases susceptibility to noise-induced hearing loss.
13-8600 was used in immunohistochemistry - paraffin section and western blot to elucidate the role of ERK isoforms to auditory function
|Kurioka T,Matsunobu T,Satoh Y,Niwa K,Endo S,Fujioka M,Shiotani A||Scientific reports (5:null)||2015|
Loss of NGF-TrkA signaling from the CNS is not sufficient to induce cognitive impairments in young adult or intermediate-aged mice.
13-8600 was used in western blot to generate and characterize mice conditionally deleted for Ngf or Trka in the central nervous system.
|Müller M,Triaca V,Besusso D,Costanzi M,Horn JM,Koudelka J,Geibel M,Cestari V,Minichiello L||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:14885)||2012|
TrkB modulates fear learning and amygdalar synaptic plasticity by specific docking sites.
13-8600 was used in western blot to study molecular pathways required for fear learning
|Musumeci G,Sciarretta C,Rodríguez-Moreno A,Al Banchaabouchi M,Negrete-Díaz V,Costanzi M,Berno V,Egorov AV,von Bohlen Und Halbach O,Cestari V,Delgado-García JM,Minichiello L||The Journal of neuroscience : the official journal of the Society for Neuroscience (29:10131)||2009|
|Not Applicable||Not Cited||
PAC1 is a direct transcription target of E2F-1 in apoptotic signaling.
13-8600 was used in western blot to demonstrate that phosphatase of activated cells 1 phosphatase is a direct transcription target of E2F-1 and mediates apoptosis
|Wu J,Jin YJ,Calaf GM,Huang WL,Yin Y||Oncogene (26:6526)||2007|
|Roscovitine targets, protein kinases and pyridoxal kinase.||Bach S,Knockaert M,Reinhardt J,Lozach O,Schmitt S,Baratte B,Koken M,Coburn SP,Tang L,Jiang T,Liang DC,Galons H,Dierick JF,Pinna LA,Meggio F,Totzke F,Schächtele C,Lerman AS,Carnero A,Wan Y,Gray N,Meijer L||The Journal of biological chemistry (280:31208)||2005|
|Regulation of SPIN90 phosphorylation and interaction with Nck by ERK and cell adhesion.||Lim CS,Kim SH,Jung JG,Kim JK,Song WK||The Journal of biological chemistry (278:52116)||2003|
Adipocyte functions are modulated by cell size change: potential involvement of an integrin/ERK signalling pathway.
13-8600 was used in western blot to test if an increase in fat cell size modulates signaling pathways by changing the relationships between the cell and the extracellular matrix.
|Farnier C,Krief S,Blache M,Diot-Dupuy F,Mory G,Ferre P,Bazin R||International journal of obesity and related metabolic disorders : journal of the International Association for the Study of Obesity (27:1178)||2003|
|Human||Not Cited||Distinct mechanisms mediate the initial and sustained phases of cell migration in epidermal growth factor receptor-overexpressing cells.||Kruger JS,Reddy KB||Molecular cancer research : MCR (1:801)||2003|
|Human||Not Cited||Alpha 7 nicotinic acetylcholine receptors mediate beta-amyloid peptide-induced tau protein phosphorylation.||Wang HY,Li W,Benedetti NJ,Lee DH||The Journal of biological chemistry (278:31547)||2003|