Catalogue# MCA-5H7: Mouse Monoclonal Antibody to Ewing Sarcoma Breakpoint Region 1 (EWSR1).
The Immunogen: Ewing sarcoma breakpoint region 1 is a protein belonging to the family of RNA binding proteins which contain a single RRM domain. It also has a single Zinc Finger domain of the ZnF_RBZ subtype found in Ran binding proteins. Ran is a small G protein related to p21-Ras which regulates the import and export of proteins to the nucleus. EWSR1 appears to be a multifunctional protein that is involved in various cellular processes, including gene expression, cell signaling, and RNA processing and transport. In particular, EWSR1 is required for cell survival in the central nervous system (1). The EWSR1 gene was identified based on its location at the chromosome 22 breakpoint of the t(11;22)(q24;q12) translocation that characterizes Ewing sarcoma and related neuroectodermal tumors. This results in the production of an aberrant fusion protein containing the N-terminus of a protein encoded by the Friend leukemia virus integration 1 (FLI1) protein and the C-terminal of the EWS protein. The chromosomal region harboring the EWS gene appears to be particularly prone to translocations, and such translocations can result in the production of several other aberrant fusion proteins, in each case with C-terminus of EWS fused with an another protein (2, see also the OMIM link). EWSR1 is ubiquitously expressed and is localized primarily in the nucleus of cell (3). It has also been found in the cytoplasm, and associated with the plasma membrane in a fashion regulated by the protein kinase Pyk2 (4). Expression of EWS in the various subcellular compartments is affected by the methylation state of its RNA-binding domain (5). Localization of EWS in different subcellular compartments reflects a dynamic distribution during cell cycle. EWS associates with and stabilizes microtubules, leading to cell cycle arrest (6). EWSR1, TAF15 and FUS/TLS are closely related proteins which are believed to have evolved by gene multiplication, and all have been implication in the etiology of some forms of Amyotrophic Lateral Sclerosis (ALS, a.k.a. Lou Gehrig's Disease) (7). Monoclonal antibody MCA-5H7 was raised against full length recombinant human EWS expressed in and purified from E. coli. The HGNC name for EWS is EWSR1.
We are OEM suppliers of this antibody- in other words we manufactured it, characterized it and generated the data presented on this page. This antibody is available from several other vendors, but we can supply it more cheaply and we can provide you with more detailed information on the properties of the antibody.
Figures: Left: Blots of whole HeLa cell lysates blotted with MCA-5H7. MCA-5H7 binds strongly and cleanly to a band at about 85 kDa. Position at which SDS-PAGE molecular weight standards run is are indicated at left. Right: An image of HeLa cell cultures stained with antibody MCA-5H7 (green) and chicken antibody to vimentin CPCA-Vim (red). Blue is a DNA stain, and it is clear that in these cells, EWS is localized along with the DNA in the nucleus.
Antibody characteristics: MCA-5H7 is a mouse IgG2b class antibody with a κ light chain. MCA-5H7 recognizes EWS specifically both in western blots and in immunocytochemical experiments. On blots MCA-5H7 reveals a prominent 85 kDa band, and on cells in tissue culture the antibody stains mainly the nucleus.
Suggestions for use: The antibody is protein G purified from tissue culture supernatant and is diluted in phosphate buffered saline preparation containing 0.05% sodium azide preservative. The antibody solution can be used at dilutions of 1:500 for immunofluorescence. For western blots try at 1:1000. A suitable control tissue is HeLa cell lysates. Avoid repeated freezing and thawing, store at 4°C or -20°C.
Omim link: press here
1: Azuma M, Embree LJ, Sabaawy H, Hickstein DD. Ewing sarcoma protein ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo. PLoS One. 2:e979 (2007).
2: Delattre O, Zucman J, Plougastel B, Desmaze C, Melot T, Peter M, Kovar H, Joubert I, de Jong P, Rouleau G, et al. Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours. Nature. 359:162-5 (1992).
3. Andersson MK, Ståhlberg A, Arvidsson Y, Olofsson A, Semb H, Stenman G, Nilsson O, Aman P. The multifunctional FUS, EWS and TAF15 proto-oncoproteins show cell type-specific expression patterns and involvement in cell spreading and stress response. BMC Cell Biol. 11:9:37 (2008).
4: Felsch JS, Lane WS, Peralta EG. Tyrosine kinase Pyk2 mediates G-protein-coupled receptor regulation of the Ewing sarcoma RNA-binding protein EWS. Curr Biol. 9:485-8 (1999).
5: Belyanskaya LL, Delattre O, Gehring H.Expression and subcellular localization of Ewing sarcoma (EWS) protein is affected by the methylation process. Exp Cell Res. Aug 15;288(2):374-81 (2003). Exp Cell Res. 288:374-81 (2003).
6: Leemann-Zakaryan RP, Pahlich S, Sedda MJ, Quero L, Grossenbacher D, Gehring H. Dynamic subcellular localization of the Ewing sarcoma proto-oncoprotein and its association with and stabilization of microtubules. J Mol Biol. 386:1-13 (2009).
7. Da Cruz S, Cleveland DW. Understanding the role of TDP-43 and FUS/TLS in ALS and beyond. Curr Opin Neurobiol. [Epub ahead of print] (2011).
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
Availability and Price: Available for shipping now, $200 US per aliquot of 100 μL of purified antibody at a concentration of 1 mg/mL, enough for hundreds of experiments. For order form press here.