Immunofluorescence analysis of Phospho-p44 MAPK + p42 MAPK pThr202 + pTyr204 Antibody was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-p44 MAPK + p42 MAPK pThr202 + pTyr204 Antibody (368800) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 45 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 cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic phosphopeptide corresponding to the region surrounding the phosphorylated Thr202 and Tyr204 residues of ERK1+2 (p44/42 MAP kinase).|
|Storage buffer||0.01M HEPES, pH 7.5, with 50% glycerol, 0.15M NaCl, 100µg/ml BSA|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:500|
|Western Blot (WB)||1:250|
* 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.
This antibody is predicted to react with Xenopus and zebrafish based on 100% sequence homology.
36-8800 is specific for the ~42k - 44k ERK/MAPK phosphorylated at Thr202 and Tyr204. Immunolabeling is blocked by the phosphopeptide used as antigen but not by the corresponding dephosphopeptide. The immunolabeling is completely eliminated by lambda-phosphatase.
Extracellular signal-regulated kinase 2 (Erk2) is also known as mitogen activated protein kinase 2, microtubule-associated protein-2 kinase MAP kinase 2, p42-MAPK or EGF receptor T669 (ERT1). Mek2 activates Erk2 by phosphorylation of neighbouring threonine-183 and tyrosine-185 residues. Erk2, a serine-threonine kinase phosphorylates microtubule associated protein-2 and myelin basic protein. This kinase is an important proximal componant of the MAP kinase pathway involved in transmitting the signals from growth factors, neurotransmitters and hormones at the cell surface to the transcriptional events in the nucleus.
Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot of Phospho-ERK1/ERK2 (Thr202, Tyr204) using Phospho-ERK1/ERK2 (Thr202, Tyr204) Rabbit Polyclonal Antibody (Product # 368800), shows increased levels of phosphorylated ERK1/ERK2 (Thr202, Tyr204) in A-431 and HeLa cell lines upon PDGF treatment. Cell Treatment validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Oncogenic Ras differentially regulates metabolism and anoikis in extracellular matrix-detached cells.
36-8800 was used in western blot to elucidate the mechanisms by which cancer cells overcome anoikis
|Mason JA,Davison-Versagli CA,Leliaert AK,Pape DJ,McCallister C,Zuo J,Durbin SM,Buchheit CL,Zhang S,Schafer ZT||Cell death and differentiation (23:1271)||2016|
The role of multicellular aggregation in the survival of ErbB2-positive breast cancer cells during extracellular matrix detachment.
36-8800 was used in western blot to study the mechanism for the metastasis of ErbB2-positive breast cancer cells.
|Rayavarapu RR,Heiden B,Pagani N,Shaw MM,Shuff S,Zhang S,Schafer ZT||The Journal of biological chemistry (290:8722)||2015|
|Human||Not Cited||The important roles of RET, VEGFR2 and the RAF/MEK/ERK pathway in cancer treatment with sorafenib.||Mao WF,Shao MH,Gao PT,Ma J,Li HJ,Li GL,Han BH,Yuan CG||Acta pharmacologica Sinica (33:1311)||2012|
|Human||Not Cited||Post-receptor IGF1 insensitivity restricted to the MAPK pathway in a Silver-Russell syndrome patient with hypomethylation at the imprinting control region on chromosome 11.||Montenegro LR,Leal AC,Coutinho DC,Valassi HP,Nishi MY,Arnhold IJ,Mendonca BB,Jorge AA||European journal of endocrinology (166:543)||2012|
|Mouse||Not Cited||Growth-factor receptor-bound protein-2 (Grb2) signaling in B cells controls lymphoid follicle organization and germinal center reaction.||Jang IK,Cronshaw DG,Xie LK,Fang G,Zhang J,Oh H,Fu YX,Gu H,Zou Y||Proceedings of the National Academy of Sciences of the United States of America (108:7926)||2011|
|Mouse||Not Cited||15-Deoxy-Delta12,14-prostaglandin J2 regulates endogenous Cot MAPK kinase kinase 1 activity induced by lipopolysaccharide.||Caivano M,Rodriguez C,Cohen P,Alemany S||The Journal of biological chemistry (278:52124)||2003|
Antihypertensive drug Valsartan promotes dendritic spine density by altering AMPA receptor trafficking.
36-8800 was used in immunohistochemistry to elucidate the molecular mechanism by which Valsartan regulates cognitive function.
|Sohn YI,Lee NJ,Chung A,Saavedra JM,Scott Turner R,Pak DT,Hoe HS||Biochemical and biophysical research communications (439:464)||2013|
|Human||Not Cited||Osteopontin is linked to p65 and MMP-9 expression in pulmonary adenocarcinoma but not in malignant pleural mesothelioma.||Frey AB,Wali A,Pass H,Lonardo F||Histopathology (50:720)||2007|
A tetra(ethylene glycol) derivative of benzothiazole aniline enhances Ras-mediated spinogenesis.
36-8800 was used in immunocytochemistry to study the effect of a novel amyloid-binding small molecule on Ras-mediated spinogenesis.
|Megill A,Lee T,DiBattista AM,Song JM,Spitzer MH,Rubinshtein M,Habib LK,Capule CC,Mayer M,Turner RS,Kirkwood A,Yang J,Pak DT,Lee HK,Hoe HS||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:9306)||2013|