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Key Documents

ABN1378

Sigma-Aldrich

Anti-Intersectin-1/ITSN1 Antibody

from rabbit

Synonyme(s) :

Intersectin-1, SH3 domain-containing protein 1A, SH3P17, Intersectin-1/ITSN1

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About This Item

Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.41

Source biologique

rabbit

Niveau de qualité

Forme d'anticorps

purified antibody

Type de produit anticorps

primary antibodies

Clone

polyclonal

Espèces réactives

rat, human

Réactivité de l'espèce (prédite par homologie)

mouse (based on 100% sequence homology), primate (based on 100% sequence homology), equine (based on 100% sequence homology), bovine (based on 100% sequence homology), feline (based on 100% sequence homology)

Technique(s)

immunoprecipitation (IP): suitable
western blot: suitable

Numéro d'accès NCBI

Numéro d'accès UniProt

Conditions d'expédition

wet ice

Modification post-traductionnelle de la cible

unmodified

Informations sur le gène

human ... ITSN1(6453)

Description générale

Intersectin-1 (UniProt Q15811; also known as Human intersectin-SH3 domain-containing protein SH3P17, SH3 domain-containing protein 1A, SH3P17, Src homology 3 domain-containing protein 1A) is encoded by the ITSN1 (also known as ITSN, SH3D1A) gene (Gene ID 6453) in human. Intersectin-1 and the related intersectin-2 are molecular scaffolds that play essential roles at the early stages of clathrin-mediated endocytosis (CME) by clustering the membrane-sculpting FCHo1/2 proteins that mark the assembly sites of clathrin-coated pits. Both ITSNs are widely expressed in tissues and share the same domain organization. Both ITSN transcripts are subjected to alternative splicing, generating short and long isoforms. The short splice form (ITSN-s) comprises two Eps15 homology (EH) domains, a coiled-coil region (CCR) and five Src 3 homology (SH3) domains. The long splice variant (ITSN-L) contains an additional Dbl homology (DH) GDP/GTP exchange domain, a plekstrin homology (PH) and C2 domains. Most of these domains mediate protein-protein interactions necessary for the assembly of multi-protein complexes. In addition to CME, ITSN1 is known to interact with multiple proteins involved in exocytosis, mitogenic signaling, actin cytoskeleton remodeling and apoptosis. ITSN1 mRNA is reported to be elevated in patients with Down syndrome and Alzheimer’s disease, and ITSN1 over-expression increases aggregation of mutant huntingtin in cultured cells.

Spécificité

This antiserum detects ITSN1, but not ITSN2. This antiserum targets EH domain sequence common to all 13 spliced variants of ITSN1 (UniProt Q15811-1 through Q15811-13). The long isoform ITSN-l (UniProt Q15811-1) is neuron-enriched and can be detected at a high level in brain. ITSN-l (~200kDa) could also be detected at low levels in some, but not all non-neuronal tissues and cell lines.

Immunogène

Epitope: EH2 domain
His-tagged recombinant protein corresponding to the EH2 domain of human Intersectin-1/ITSN1.

Application

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated Ruk/CIN85 with transiently expressed Omni-tagged ITSN1 in an EGF stimulation-independent manner using a stable MCF-7 culture overexpressing Ruk/CIN85 (Novokhatska, O., et al. (2009). Cell Signal. 21(5):753-759).
Immunoprecipitation Analysis: A representative lot co-immunoprecipitated ITSN2 with ITSN1 from HEK293 cells (Novokhatska, O., et al. (2013). PLoS One. 8(7):e70546).
Immunoprecipitation Analysis: A representative lot immunoprecipitated ITSN1 in lysates prepared from growing HEK293, HeLa, MCF-7, and MDA-MB-231 cultures (Novokhatska, O., et al. (2013). PLoS One. 8(7):e70546).
Western Blotting Analysis: A representative lot detected ITSN1, but not ITSN2, by Western blotting using total HEK293 cell lysate or ITSN1 immunoprecipitate obtained from HeLa cell lysates (Novokhatska, O., et al. (2013). PLoS One. 8(7):e70546).
This Anti-Intersectin-1/ITSN1 Antibody is validated for use in Western Blotting and Immunoprecipitation for the detection of Intersectin-1/ITSN1.

Qualité

Evaluated by Western Blotting in rat brain cytosol tissue lysate.

Western Blotting Analysis: Western Blotting Analysis: 0.5 µg/mL of this antibody detected Intersectin-1/ITSN1 in 10 µg of rat brain cytosolic preparation.

Description de la cible

~160 kDa observed. Target band size(s) may vary depending on the isoform(s) present in the sample(s).

Forme physique

Format: Purified

Autres remarques

Concentration: Please refer to lot specific datasheet.

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Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 1

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


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Consulter la Bibliothèque de documents

Claudia Puri et al.
Developmental cell, 53(2), 154-168 (2020-04-22)
Autophagy involves engulfment of cytoplasmic contents by double-membraned autophagosomes, which ultimately fuse with lysosomes to enable degradation of their substrates. We recently proposed that the tubular-vesicular recycling endosome membranes were a core platform on which the critical early events of

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