Application

This Anti-Intersectin-1/ITSN1 Antibody is validated for use in Western Blotting and Immunoprecipitation for the detection of Intersectin-1/ITSN1.

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).

General description

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.

Immunogen

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

Epitope: EH2 domain

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Format: Purified

Quality

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.

Specificity

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.

Target description

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