ITSN1 Antibody, FITC conjugated
Description
Definition and Purpose
The ITSN1 Antibody, FITC conjugated is a fluorescently tagged antibody designed to detect ITSN1 protein in biological samples. FITC (fluorescein isothiocyanate) is a green fluorescent dye excitable at 485–495 nm and emits at 515–545 nm, making it ideal for imaging techniques like immunofluorescence (IF) and flow cytometry .
Primary Uses
FITC Advantages
Key Studies Using FITC-Conjugated ITSN1 Antibodies
Select FITC-Conjugated ITSN1 Antibodies
Note: FITC-conjugated variants may require direct supplier confirmation for availability.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- WIP enhances the interaction of N-WASP with ITSN1 and promotes ITSN1/beta-actin association. PMID: 29958948
- Intersectin-s binds DENND2B, a guanine nucleotide exchange factor for the exocytic GTPase Rab13, and this interaction promotes recycling of ligand-free EGFR to the cell surface. This study reveals a novel mechanism controlling the fate of internalized EGFR with important implications for cancer. PMID: 29030480
- This study reports that vaccinia virus A36 contains three NPF motifs that interact with the Eps15 homology (EH) domains in intersectin-1 and Eps15 to promote the release of virus from infected cells and their subsequent spread. PMID: 27670116
- Silencing ITSN1 in neuroblastoma cells led to decreased tumor growth in an orthotopic mouse model. Orthotopic animal models can provide insight into the role of ITSN1 pathways in neuroblastoma tumorigenesis. PMID: 28787396
- These findings provide the basis for further functional investigations of the ITSN/CR16 complex that may play an important role in actin remodeling and cellular invasion. PMID: 28161632
- Abnormal expression of the intersectin1-L protein in epileptic brain tissue may play an important role in epilepsy, especially refractory epilepsy. PMID: 25783631
- This study suggests critical roles of ITSN1-S in malignant glioma proliferation, indicating a potential usage of ITSN1-S in the therapeutic intervention as a novel molecular target. PMID: 25832561
- Our observations suggest that ITSN1 is an important general regulator of Cdc42-, Nck- and N-WASP-dependent actin polymerisation. PMID: 24284073
- ITSN1 and ITSN2 bind similar proline-rich ligands but are differentially recognized by SH2 domain-containing proteins. PMID: 23936226
- Silencing ITSN1 significantly inhibits the anchorage independent growth of tumor cells in vitro. PMID: 22266851
- The neuron-specific isoform of the stable tubule-only polypeptide (STOP) interacts with SH3A domain of ITSN1. PMID: 22750298
- ITSN-1s, via its SH3A domain, has the unique ability to regulate dyn2 assembly-disassembly and function during endocytosis. PMID: 21129155
- ITSN1 has two isoforms: ITSN1-long and ITSN1-short. siRNA-mediated down regulation of ITSN1-short inhibits migration and invasion of glioma cells. PMID: 21503949
- This study reveals a link between overexpression of specific ITSN1 isoforms and behavioral phenotypes. PMID: 21876463
- This novel mammalian ITSN1 isoform possesses a significantly altered domain structure and performs specific protein-protein interactions. PMID: 21712076
- Downregulation of intersectin-1s induces apoptosis of brain glioblastoma cells. PMID: 20493827
- This study investigated the inhibitory mechanism of ITSN1L and identified a novel short amino acid motif that mediates autoinhibition. This motif is located in the SH3 DH linker region, and the study shows that W1221 acts as a key residue in establishing inhibitory interaction. PMID: 20842712
- These findings expand the role of ITSN1 as a scaffolding molecule bringing together components of endocytic complexes. PMID: 20946875
- The identification of fifteen novel transcriptional isoforms of the human ITSN1 gene with full-length coding sequences is reported. PMID: 19777371
- Microexons provide a mechanism for the control of tissue-specific interactions of ITSN1 and Src with their partners. PMID: 20659428
- Intersectin interaction with dynamin was important in regulating the fission and internalization of caveolae. PMID: 12960435
- This review discusses the roles of ITSN1 and DSCR1 in Down syndrome, Alzheimer's disease, endocytosis and vesicle trafficking. PMID: 16442855
- These results extend the current role of intersectin-1L in endocytosis to a function in exocytosis and support the idea that intersectin-1L is an adaptor that coordinates exo-endocytotic membrane trafficking in secretory cells. PMID: 16874303
- ITSN forms a complex with Cbl in vivo mediated by the Src homology (SH) 3 domains binding to the Pro-rich COOH terminus of Cbl. This interaction stimulates the ubiquitylation and degradation of the activated EGFR. PMID: 16914641
- These findings demonstrate a novel role of ITSN-1s as a negative regulator of the mitochondrial pathway-dependent apoptosis secondary to activation of the Erk1/2 survival signaling pathway. PMID: 17405881
- These results demonstrate that alternative splicing leads to the formation of two pools of ITSN1 with potentially different properties in neurons, affecting ITSN1 function as an adaptor protein. PMID: 18539136
- These data provide a molecular link between SHIP2 and ITSN1 which are involved in receptor endocytosis regulation. PMID: 18692052
- Intersectin 1 forms a complex with adaptor protein (SH3KBP1 protein), implicated in downregulation of receptor tyrosine kinases. PMID: 19166927
- The higher exchange activity of ITSN1L towards the GDP-bound conformation of Cdc42 could represent an evolutionary adaptation of this GEF that ensures nucleotide exchange towards the formation of the signaling-active GTP-bound form of Cdc42. PMID: 19356586
Q&A
What experimental validations are required to confirm FITC-conjugated ITSN1 antibody specificity in breast cancer models?
To ensure antibody specificity, researchers should employ a multi-modal validation strategy:
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Knockout Controls: Use ITSN1 gene-edited cell lines (e.g., CRISPR/Cas9 knockout) to verify signal absence in Western blot (WB) and immunofluorescence (IF) .
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Cross-Reactivity Profiling: Test antibody reactivity against homologous proteins (e.g., ITSN2) using lysates from ITSN1-deficient and ITSN2-overexpressing systems .
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Epitope Mapping: Compare binding performance against antibodies targeting distinct ITSN1 domains (e.g., N-terminal vs. SH3 domains) .
Table 1: Validation Strategies Across Studies
How can researchers optimize FITC-conjugated ITSN1 antibody dilution for fluorescence microscopy?
Optimal dilution depends on fixation methods and target abundance:
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Fixation: For formaldehyde-fixed cells, use 1:200–1:500 dilution to mitigate epitope masking .
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Signal-to-Noise Ratio: Pre-block with 5% BSA + 0.1% Triton X-100 to reduce non-specific binding .
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Counterstaining Compatibility: Pair with far-red nuclear markers (e.g., DRAQ5) to avoid spectral overlap .
How can FITC-conjugated ITSN1 antibodies resolve contradictory localization data in breast cancer subtypes?
Discrepancies in ITSN1 nuclear vs. cytoplasmic localization arise from:
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Post-Translational Modifications: Phosphorylation at Ser⁷⁷⁶ alters subcellular distribution .
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Tumor Subtype Context: Luminal subtypes show predominant nuclear ITSN1-S, while basal-like cancers exhibit cytoplasmic retention .
Methodological Recommendations:
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Perform subcellular fractionation followed by WB to quantify compartment-specific ITSN1 levels .
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Use proximity ligation assays with NDH II/PI3KC2α antibodies to map interaction niches .
What advanced multiplexing strategies incorporate FITC-conjugated ITSN1 antibodies to study endocytic-kinase crosstalk?
Combine FITC-ITSN1 with:
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Phospho-Specific AKT Antibodies (Cy5): To correlate ITSN1-PI3KC2α interactions with AKT activation .
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EGFR-TRITC: Track ITSN1’s role in EGFR endocytosis using live-cell imaging .
Table 2: Multiplex Assay Design
How can super-resolution microscopy enhance FITC-conjugated ITSN1 antibody applications?
dSTORM (direct Stochastic Optical Reconstruction Microscopy) achieves 20 nm resolution to:
Critical Parameters:
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Use glucose oxidase-based imaging buffers to maintain fluorophore stability.
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Acquire ≥10,000 frames per dataset to ensure sufficient localization events .
How to address false-positive ITSN1 signals in immunohistochemistry (IHC)?
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Antigen Retrieval: Optimize citrate buffer pH (6.0 vs. 8.0) based on tissue fixation duration .
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Endogenous Fluorescence Quenching: Treat sections with 0.3% Sudan Black B to eliminate lipofuscin autofluorescence .
