Key Spec Table
|Species Reactivity||Key Applications||Host||Format||Antibody Type|
|H, M, R||IP, WB||M||Purified||Monoclonal Antibody|
|Description||Anti-SETA/CIN85/Ruk/SH3KBPI Antibody, clone 179.1.E1|
|Presentation||of 0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%|
|Application||This Anti-SETA/CIN85/Ruk/SH3KBPI Antibody, clone 179.1.E1 is validated for use in IP, WB for the detection of SETA/CIN85/Ruk/SH3KBPI.|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||2 years at -20°C|
|Material Size||200 µg|
|Anti-SETA/CIN85/Ruk/SH3KBPI, clone 179.1.E1 - 24641||24641|
|Anti-SETA/CIN85/Ruk/SH3KBPI, clone 179.1.E1 -2783096||2783096|
|Reference overview||Application||Pub Med ID|
|CIN85 modulates the down-regulation of Fc gammaRIIa expression and function by c-Cbl in a PKC-dependent manner in human neutrophils.|
Marois, L; Vaillancourt, M; Paré, G; Gagné, V; Fernandes, MJ; Rollet-Labelle, E; Naccache, PH
The Journal of biological chemistry 286 15073-84 2011
We previously described a non-classical mechanism that arrests FcγRIIa signaling in human neutrophils once engaged by immune complexes or opsonized pathogens. The engagement of FcγRIIa leads to its ubiquitination by the ubiquitin ligase c-Cbl and degradation by the proteasome. Herein, we further examined some of the events regulating this novel pathway. The adaptor protein CIN85 was described in other systems to be involved in the regulation of the c-Cbl-dependent pathway. We found that CIN85 is expressed in human neutrophils and that it translocates like c-Cbl from the cytosol to the plasma membrane following receptor cross-linking. CIN85 was also recruited to the same subset of high density detergent-resistant membrane fractions in which stimulated FcγRIIa partitioned with c-Cbl. The integrity of these microdomains is essential to the FcγRIIa degradation process because the cholesterol-depleting agent methyl-β-cyclodextrin inhibits this event. Silencing the expression of CIN85 by siRNA in dibutyryl cyclic AMP-differentiated PLB 985 cells prevented FcγRIIa degradation and increased IgG-mediated phagocytosis. Confocal microscopy revealed that the presence of CIN85 is essential to the proper sorting of FcγRIIa during endocytosis. We also provide direct evidence that CIN85 is a substrate of serine/threonine kinase PKCs. Classical PKCs positively regulate FcγRIIa ubiquitination and degradation because these events were inhibited by Gö6976, a classical PKC inhibitor. We conclude that the ubiquitination and degradation of stimulated FcγRIIa mediated by c-Cbl are positively regulated by the adaptor protein CIN85 in a PKC-dependent manner and that these events contribute to the termination of FcγRIIa signaling.
|Studying protein isoforms of the adaptor SETA/CIN85/Ruk with monoclonal antibodies.|
Finniss, Susan, et al.
Biochem. Biophys. Res. Commun., 325: 174-82 (2004) 2004
SETA/CIN85/Ruk is a multifunctional adaptor protein involved in signal transduction and attenuation downstream of receptor tyrosine kinases. It has a modular structure, and various isoforms that combine different protein-protein interaction domains have been proposed based on cDNA analysis. As a first step towards understanding SETA/CIN85/Ruk isoforms at the protein level, we have characterized 5 monoclonal antibodies against this protein. Three of these were used to study lysates fractionated on a pH gradient, leading to the identification of various SETA/CIN85/Ruk proteins on the basis of pI and apparent molecular weight. While good correspondence with proteins predicted from cDNA analysis was found for two isoforms, in most cases it was not possible to make an unequivocal assignment. We conclude that additional splice variants remain to be described, and that a deeper understanding of SETA/CIN85/Ruk post-translational processing and modification is necessary to gain further understanding of this complex gene product.
|SETA: a novel SH3 domain-containing adapter molecule associated with malignancy in astrocytes.|
Bögler, O, et al.
Neuro-oncology, 2: 6-15 (2000) 2000
|The glioma-associated protein SETA interacts with AIP1/Alix and ALG-2 and modulates apoptosis in astrocytes.|
Chen, B, et al.
J. Biol. Chem., 275: 19275-81 (2000) 2000
Expression of the src homology 3 (SH3) domain-containing expressed in tumorigenic astrocytes (SETA) gene is associated with the tumorigenic state in astrocytes. SETA encodes a variety of adapter proteins containing either one or two SH3 domains, as suggested by the sequence heterogeneity of isolated cDNAs. Using both SH3 domains in a yeast two-hybrid screen of a glial progenitor cell cDNA library, we isolated the rat homolog of the ALG-2-interacting protein 1 or ALG-2-interacting protein X (AIP1/Alix). In vitro confrontation experiments showed that the SH3-N domain of SETA interacted with the proline-rich C terminus of AIP1. In co-immunoprecipitation experiments, SETA and AIP1 interacted and could form a complex with apoptosis-linked gene 2 protein. Endogenous SETA and AIP1 proteins showed similar patterns of staining in primary rat astrocytes. Misexpression of a variety of SETA protein isoforms in these astrocytes revealed that they localized to the actin cytoskeleton. Furthermore, SETA proteins containing the SH3-N domain were able to sensitize astrocytes to apoptosis induced by UV irradiation. Expression of the isolated SH3-N domain had the greatest effect in these experiments, indicating that interference in the interaction between endogenous SETA and AIP1 sensitizes astrocytes to apoptosis in response to DNA damage.