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07-1364
Sigma-AldrichAnti-ECT2 Antibody
Anti-ECT2, Cat. No. 07-1364, is a rabbit polyclonal antibody that detects ECT2 protein and is tested for use in Immunocytochemistry and Western Blotting.
More>>Anti-ECT2, Cat. No. 07-1364, is a rabbit polyclonal antibody that detects ECT2 protein and is tested for use in Immunocytochemistry and Western Blotting. Less<<
Anti-ECT2 Antibody: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
Protein ECT2 (UniProt: Q9H8V3; also known as Epithelial cell-transforming sequence 2 oncogene) is encoded by the ECT2 gene (Gene ID: 1894) in human. ECT2 is a Guanine nucleotide exchange factor (GEF) that catalyzes the exchange of GDP for GTP and promotes guanine nucleotide exchange on the Rho family members of small GTPases. It is expressed in lung epithelial cells. Its expression is also observed in squamous cell carcinoma, primary non-small cell lung cancer tumors and lung adenocarcinoma. Its levels are up-regulated by calcium in cells forming cell-cell contact sites. It is also up-regulated by DNA damaging agents like hydrogen peroxide and ionizing radiation. ECT2 is required for signal transduction pathways involved in the regulation of cytokinesis. It plays a role in the control of mitotic spindle assembly and regulates the activation of Cdc42 in metaphase for the process of spindle fibers attachment to kinetochores before chromosome congression. It contains two BCRT domains (aa 171-260 and 266-354) and the second BCRT domain is involved in inhibition, probably by helping to hamper RhoA binding. ECT2 is autoinhibited by the C-terminal PH domain (aa 675-794) that folds back and binds to the surface of the DH domain (aa 452-641), blocking binding of RhoA to the catalytic center of the DH domain. ECT2 is phosphorylated by PLK1 in vitro and is hyperphosphorylated during the G2 phase of the cell cycle. Phosphorylation at threonine 373 occurs during the G2/M phase that relieves its auto-inhibition status and stimulates its GEF activity. Phosphorylation at threonine 444 in G2/M phase is required for subsequent binding with PLK1 and Rho exchange activation. (Ref.: Chen, M., et al. (2020). Proc. Natl. Acad. Sci. USA. 117(2); 1027-1035; Kim, JE., et al. (2005). J. Biol. Chem. 280(7); 5733-5739; Tatsumoto, T., et al. (1999). J. Cell Biol. 147(5); 921-928).
References
Product Information
Format
Affinity Purified
Control
A549 cell lysate
Presentation
Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide
Anti-ECT2, Cat. No. 07-1364, is a rabbit polyclonal antibody that detects ECT2 protein and is tested for use in Immunocytochemistry and Western Blotting.
Key Applications
Western Blotting
Immunocytochemistry
Application Notes
Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Protein ECT2 in A549 cells.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biological Information
Immunogen
KLH-conjugated linear peptide corresponding to 15 amino acids from the C-terminal region of human ECT2.
Epitope
C-terminus
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Rabbit
Specificity
This rabbit polyclonal antibody detects ECT2. It targets an epitope within 15 amino acids from the C-terminal region.
Species Reactivity
Human
Species Reactivity Note
Human. Predicted to react with Mouse, Monkey, Bovine, Opossum, Porcine, Rat based on 100% sequence homology.
FUNCTION: Binds highly specifically to RhoA, RhoC and Rac proteins, but does not appear to catalyze guanine nucleotide exchange By similarity. SUBUNIT STRUCTURE: Associates with RACGAP1 at anaphase and during cytokinesis. SEQUENCE SIMILARITES: Contains 2 BRCT domains. Contains 1 DH (DBL-homology) domain. Contains 1 PH domain. CAUTION: It is uncertain whether Met-1 or Met-146 is the initiator.
Molecular Weight
~105 kDa observed; 103.51 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in A549 cell lysate.
Western Blotting Analysis: A 1:2,000 dilution of this antibody detected Protein ECT2 in A549 cell lysate.
Usage Statement
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.
MgcRacGAP interacts with cingulin and paracingulin to regulate Rac1 activation and development of the tight junction barrier during epithelial junction assembly. Guillemot, L; Guerrera, D; Spadaro, D; Tapia, R; Jond, L; Citi, S Molecular biology of the cell
25
1995-2005
2014
The regulation of Rho-family GTPases is crucial to direct the formation of cell-cell junctions and tissue barriers. Cingulin (CGN) and paracingulin (CGNL1) control RhoA activation in epithelial cells by interacting with RhoA guanidine exchange factors. CGNL1 depletion also inhibits Rac1 activation during junction assembly. Here we show that, unexpectedly, Madin-Darby canine kidney epithelial cells depleted of both CGN and CGNL1 (double-KD cells) display normal Rac1 activation and tight junction (TJ) formation, despite decreased junctional recruitment of the Rac1 activator Tiam1. The expression of the Rac1 inhibitor MgcRacGAP is decreased in double-KD cells, and the barrier development and Rac1 activation phenotypes are rescued by exogenous expression of MgcRacGAP. MgcRacGAP colocalizes with CGN and CGNL1 at TJs and forms a complex and interacts directly in vitro with CGN and CGNL1. Depletion of either CGN or CGNL1 in epithelial cells results in decreased junctional localization of MgcRacGAP but not of ECT2, a centralspindlin-interacting Rho GEF. These results provide new insight into coordination of Rho-family GTPase activities at junctions, since apical accumulation of CGN and CGNL1 at TJs during junction maturation provides a mechanism to spatially restrict down-regulation of Rac1 activation through the recruitment of MgcRacGAP.
