UNC119 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research indicates that UNC119 is a regulator of the RASSF6-MDM2-p53 axis and functions as a tumor suppressor. PMID: 29931788
- This review highlights the trafficking pathways required for constructing the cilium, emphasizing UNC119's role and the intricacies involved in ciliary trafficking. While UNC119 is essential for ciliary functions, it also interacts with non-ciliary proteins, impacting other cellular processes. PMID: 28935136
- Data demonstrate that the solubilising factor UNC119 sequesters myristoylated Src family protein tyrosine kinases (SFKs) to maintain its enrichment at the plasma membrane, facilitating signal transduction. PMID: 28740133
- Studies indicate that UNC119 and PDE6D bind to the lipid-modified ciliary cargo and the specific release of the cargo in the cilia by the ciliary small G-protein Arl3 in a GTP-dependent manner. PMID: 27911709
- Novel biochemical and structural insights into the interaction of myristoylated cargo with Unc119 protein and their release by Arl2/3 have been reported. PMID: 27481943
- UNC119a acts as a bridge, transmitting Fyn signals to Rab11, leading to the completion of cytokinesis. PMID: 23535298
- The subcellular distribution of UNC119a remains largely unchanged under various illumination conditions, correlating with the profile of Galpha(t1) following its light-dependent translocation. PMID: 23072788
- The discovery of the UNC119 defect provides a molecular mechanism for a subset of patients with previously unexplained disease. This review summarizes recent findings on the UNC119 mutation in ICL. PMID: 22729960
- Crystal structures of Arl3 in complex with UNC119a reveal the molecular basis of specificity. The N-terminal amphipathic helix of Arl3.GppNHp remains bound on the surface of the protein, despite not being displaced by the interswitch toggle. PMID: 22960633
- The interaction of transducin with uncoordinated 119 protein (UNC119) has implications for the model of transducin trafficking in rod photoreceptors. PMID: 21712387
- This study demonstrates that UNC119 is a Galpha subunit cofactor essential for G protein trafficking in sensory cilia. PMID: 21642972
- Increased expression of Unc119 promotes T helper type (Th)2 cells, inhibits Th1 cell differentiation, and contributes to the pathogenesis of asthma in humans. PMID: 20220094
- UNC119 has been identified as an activator of SRC-type tyrosine kinases. PMID: 12496276
- The presence of ARL2 in the retina and its co-localization with HRG4 has been confirmed. Amino acid residues of PDEdelta involved in binding ARL2 and forming a hydrophobic pocket have been shown to be highly conserved in HRG4. PMID: 12527357
- Unc119 plays a significant role in fibrotic processes through myofibroblast differentiation. Unc119 increases the kinase activity of Fyn and associates with it in coprecipitation and colocalization studies. PMID: 17579091
- Unc119 orchestrates the recruitment of the actin-based motor protein, myosin 5B, and the organization of multiprotein complexes on endosomes. The Unc119-regulated pathway is essential for immunological synapse formation and T cell activation. PMID: 19592652
- Research demonstrates a role for Unc119 in clathrin- and caveolae-based endocytosis, as well as macropinocytosis. Depletion of Unc119 in fibroblasts increases the uptake of FM4-64, albumin, viruses, and ligand-coated beads. PMID: 19781630
Q&A
What are the primary applications of UNC119 antibodies in neurobiology and immunology research?
UNC119 antibodies are widely used to investigate synaptic protein trafficking, T-cell receptor (TCR) signaling, and lipid-binding chaperone functions. Key methodologies include:
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Western Blot: Detecting UNC119 expression in retinal photoreceptors (27 kDa band) and T-cell lysates .
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Immunohistochemistry (IHC): Localizing UNC119 in cytoplasmic compartments of photoreceptor inner segments and synaptic terminals .
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Immunofluorescence/Immunocytochemistry (IF/ICC): Visualizing UNC119 redistribution during TCR activation or synaptic vesicle cycling .
Recent studies have identified discrepancies in UNC119 localization patterns between retinal neurons (synaptic terminals) and T cells (cytosolic pools) . Researchers must validate antibody specificity using knockout controls (e.g., UNC119−/− mice) .
How to select appropriate UNC119 antibodies for specific experimental designs?
Critical parameters for antibody selection:
For T-cell studies, prioritize antibodies validated in activation assays (e.g., co-precipitation with CD3ζ/Lck) .
What controls are necessary when validating UNC119 antibody specificity?
A three-tiered validation approach is recommended:
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Genetic Controls: Compare wild-type and UNC119−/− tissues via Western blot . Example: Retinal lysates from UNC119−/− mice show no 27 kDa band .
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Competition Assays: Pre-incubate antibodies with recombinant UNC119 protein (1–2 µg/mL) to confirm signal loss .
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Subcellular Fractionation: Confirm cytoplasmic localization in resting T cells versus membrane recruitment post-TCR activation .
Failure to implement these controls may lead to misinterpretation of UNC119’s role in synaptic vesicle recycling or LCK kinase trafficking .
How does UNC119 regulate LCK/FYN kinase activity in TCR signaling, and how can this be experimentally dissected?
UNC119 enhances Src-family kinase activation through two mechanisms:
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Direct Binding: Co-precipitation assays show UNC119 interacts with LCK’s SH3 domain (Kd ≈ 120 nM) .
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Spatial Regulation: Live-cell imaging reveals UNC119 mediates LCK translocation to membrane lipid rafts upon CD3/CD28 co-stimulation .
Methodological Pipeline:
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Kinase Activity Assays: Measure Y394-LCK phosphorylation in UNC119-inhibited T cells (squarunkin A, 10 µM) .
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Proximity Ligation Assay (PLA): Quantify UNC119-LCK interactions in Jurkat cells stimulated with anti-CD3/CD28 beads .
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Single-Cell Phosphoproteomics: Use mass cytometry (CyTOF) to profile ZAP70/SLP76 phosphorylation in UNC119-depleted CD8+ T cells .
Recent findings indicate UNC119 inhibition reduces ZAP70 phosphorylation by 63% (±8%) in primary T cells, mimicking LCK deficiency .
How to resolve conflicting data on UNC119’s role in synaptic transmission versus immune function?
Discrepancies arise from tissue-specific isoforms and experimental models:
Resolution Strategy:
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Isoform-Specific Knockdown: Use siRNA targeting UNC119A (retina-enriched) vs. UNC119B (immune-enriched) .
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Contextualized Stimulation: Compare light-adapted retinal slices (500 Rh*/rod) versus anti-CD3/CD28-activated T cells .
What advanced techniques quantify UNC119-dependent protein trafficking in live cells?
For dynamic studies, employ HaloTag-UNC119 fusions paired with JF646 fluorophores (t1/2 ≈ 28 min for synaptic turnover) .
How does UNC119 influence the RASSF6-MDM2-p53 axis in cancer models?
UNC119 stabilizes p53 by enhancing RASSF6-MDM2 binding (Kd improvement from 210 nM to 89 nM) . Experimental approaches:
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Co-Immunoprecipitation: Use HCT116 cells treated with 5 µM nutlin-3 to disrupt MDM2-p53 binding .
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DNA Repair Assays: GammaH2AX foci quantification shows UNC119 depletion delays repair (48% foci at 6 hr vs. 22% in controls) .
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Ploidy Analysis: FACS reveals 14.3% polyploid cells in UNC119-depleted lines post-doxorubicin .
Clinical correlation: TCGA data link low UNC119 expression to reduced survival in colorectal adenocarcinoma (HR = 1.84, p = 0.009) .
How to address non-specific staining in UNC119 IHC?
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Preabsorption Test: Incubate antibody with 5x molar excess UNC119 peptide (aa 45-60) for 1 hr .
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Fixation Optimization: Paraformaldehyde (4%) over methanol preserves UNC119 epitopes in retinal sections .
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Signal Confirmation: Compare staining patterns across monoclonal (e.g., clone 7A3) and polyclonal antibodies .
A 2025 study found 23% of commercial UNC119 antibodies cross-react with UNC119B; always verify isoform specificity .
Why do UNC119 inhibition studies yield variable outcomes across T-ALL subtypes?
Variability stems from differential LCK dependency:
| T-ALL Subtype | UNC119i Efficacy (IC50) | LCK Expression |
|---|---|---|
| CCRF-CEM | 1.2 µM | High (+++) |
| MOLT-4 | 8.7 µM | Low (+) |
| PDX #12 | 0.9 µM | Intermediate (++) |
Solution: Pre-screen patient-derived xenografts (PDX) for LCK-Y394 phosphorylation via Phosflow . Combine UNC119i with dexamethasone to overcome glucocorticoid resistance .
What computational tools predict UNC119-binding partners in novel contexts?
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AlphaFold-Multimer: Models UNC119-myristoylated protein interactions (e.g., ARL3, PDE6D) .
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Kinase-Ligand Docking: HADDOCK simulations identify UNC119 residues critical for LCK-SH3 binding (Asp109, Glu112) .
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ScRNA-Seq Meta-Analysis: Reanalyze datasets (GSE145197) to correlate UNC119 expression with T-cell exhaustion markers (LAG3, TOX) .
Experimental validation via BioID proximity labeling is recommended for high-confidence interactors .
