Customize Unc13c Antibody

Description

Introduction to UNC13C Antibody

The UNC13C antibody is a specialized immunological reagent designed to detect and study the UNC13C protein (Unc-13 Homolog C), encoded by the UNC13C gene in humans. This protein, also known as Munc13-3, is a member of the UNC13 family, which regulates synaptic vesicle priming and exocytosis in neurons and other secretory cells . Beyond its neurological role, UNC13C has emerged as a critical player in cancer biology, with implications in oral squamous cell carcinoma (OSCC) and hepatocellular carcinoma (HCC) . The antibody is widely used in research to investigate UNC13C's expression patterns, functional mechanisms, and clinical relevance.

Molecular and Functional Characteristics of UNC13C

UNC13C is a 250.9 kDa protein containing conserved C1 and C2 domains, enabling interactions with calcium ( $\text{Ca}^{2+}$ ) and diacylglycerol (DAG) . These domains facilitate its role in membrane fusion events during neurotransmitter release and immune cell cytotoxicity .

Key Features of UNC13C:

Gene Location: Human UNC13C (Chromosome 15q21.1) .
Protein Domains: C1 (DAG-binding), C2 ( $\text{Ca}^{2+}$ -binding), and a central helical region .
Cellular Roles: Synaptic vesicle priming, exocytosis, and regulation of epithelial-mesenchymal transition (EMT) in cancer .

Role in Oral Squamous Cell Carcinoma (OSCC)

UNC13C acts as a tumor suppressor in OSCC:

Expression: Downregulated in OSCC tissues compared to normal margins .
Functional Impact: Overexpression inhibits migration and invasion of SCC-9 and SAS cell lines by suppressing EMT markers (e.g., Vimentin, Slug, Snail) and upregulating tight junction proteins (Claudin-1) .
Prognostic Value: Low UNC13C correlates with advanced clinical stage and poor survival ( $p = 0.0231$ ) .

Table 1: UNC13C Expression in OSCC Clinical Outcomes

Parameter	Low UNC13C vs. High UNC13C	P-Value
5-Year Survival Rate	45% vs. 72%	0.0231
Metastasis Incidence	68% vs. 32%	<0.001

Implications in Hepatocellular Carcinoma (HCC)

In HCC, UNC13C exhibits paradoxical roles:

Expression: High cytoplasmic UNC13C correlates with advanced T stage, AJCC stage, and reduced survival .
Prognostic Marker: Multivariate analysis identifies UNC13C as an independent prognostic factor (HR = 1.4, $p = 0.042$ ) .

Table 2: Cox Regression Analysis of UNC13C in HCC Survival

Variable	Hazard Ratio (HR)	95% CI	P-Value
Low UNC13C Expression	1.0 (Reference)	—	—
High UNC13C Expression	1.4	1.011–1.822	0.042*
Lymph Node Metastasis	4.3	1.818–10.064	0.001**

Clinical and Therapeutic Implications

Diagnostic Utility:
- UNC13C immunohistochemistry (IHC) is used to assess protein levels in tumor tissues (Antibody: Sigma-Aldrich HPA041516, 1:20–1:50 dilution) .
- Low UNC13C in OSCC predicts poor prognosis, while high levels in HCC indicate aggressive disease .
Therapeutic Potential:
- Restoring UNC13C function may inhibit metastasis in OSCC by reversing EMT .
- In HCC, targeting UNC13C could improve survival outcomes, though its oncogenic role requires further study .

Future Research Directions

Mechanistic Studies: Elucidate UNC13C’s dual roles in cancer progression and synaptic function .
Therapeutic Development: Explore small-molecule activators (for OSCC) or inhibitors (for HCC) targeting UNC13C pathways .
Biomarker Validation: Conduct multi-center studies to standardize UNC13C IHC protocols for clinical use .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

Unc13c antibody; Unc13h3 antibody; Protein unc-13 homolog C antibody; Munc13-3 antibody

Target Names

Unc13c

Uniprot No.

Q62770

Target Background

Function

Unc13c antibody targets a protein that may play a crucial role in vesicle maturation during the process of exocytosis. This protein is a potential target for diacylglycerol, a second messenger signaling molecule. Additionally, Unc13c may be involved in regulating synaptic transmission at parallel fiber - Purkinje cell synapses.

Database Links

KEGG: rno:286931

STRING: 10116.ENSRNOP00000010330

UniGene: Rn.11278

Protein Families

Unc-13 family

Subcellular Location

Cytoplasm. Membrane; Peripheral membrane protein. Cell junction, synapse, presynaptic cell membrane; Peripheral membrane protein.

Tissue Specificity

Exclusively expressed in brain, predominantly in the cerebellum.

Q&A

Basic Research Questions

How to validate UNC13C antibody specificity in Western blotting for oral squamous cell carcinoma (OSCC) models?

Method: Perform siRNA-mediated UNC13C knockdown in OSCC cell lines (e.g., SCC-9, SAS) and compare protein expression levels via Western blot. Use β-actin as a loading control .
Controls: Include lysates from non-transfected cells and cells transfected with scrambled siRNA.
Troubleshooting: Non-specific bands may arise due to cross-reactivity with homologous proteins (e.g., UNC13A/B). Optimize blocking conditions (5% BSA in TBST) and antibody dilution (1:1,000–1:2,000) .

How to select an appropriate UNC13C antibody for flow cytometry?

Criteria:
- Epitope location: Prefer antibodies targeting extracellular domains (e.g., aa 1838–1888) .
- Cross-reactivity: Verify reactivity with human samples using recombinant protein controls .
- Conjugation: Use FITC/PE-conjugated antibodies for surface staining; validate with isotype controls .

Advanced Research Questions

How to design experiments investigating UNC13C’s role in EMT signaling?

Experimental workflow:
- Overexpression/knockdown: Transfect OSCC cells with pUNC13C-Myc or siRNA .
- Phenotypic assays: Quantify migration (Transwell) and invasion (Matrigel-coated Transwell) post-transfection .
- Molecular profiling: Perform Western blotting for EMT markers (E-cadherin↓, Vimentin↑, Twist↓) and RNA-seq to identify downstream targets .
Data interpretation: Use multivariate analysis to link UNC13C expression levels with clinical stage and metastasis risk .

How to resolve contradictions in UNC13C antibody performance across studies?

Common issues:

Issue	Solution
Variable IHC staining in FFPE tissues	Optimize antigen retrieval (citrate buffer, pH 6.0) and validate with positive/negative controls .
Discrepant Western blot bands	Validate via CRISPR knockout lines and epitope mapping .
Inconsistent functional data	Standardize assay conditions (e.g., serum concentration in migration assays) .

What methodologies are suitable for studying UNC13C-antibody interactions in therapeutic contexts?

In vitro:
- Antibody-drug conjugates (ADCs): Conjugate UNC13C antibodies to toxins (e.g., MMAE) and test cytotoxicity in 3D tumor spheroids .
- Humanization: Engineer chimeric antibodies using CDR grafting (e.g., replace murine FR residues with human counterparts) .
In vivo: Use PDX models to evaluate tumor regression post-ADC administration .

Data Tables

Table 1. UNC13C Modulation Effects on EMT Markers in OSCC

Marker	Change with UNC13C Overexpression	Functional Impact
E-cadherin	↓	Reduced cell-cell adhesion
Vimentin	↓	Inhibited mesenchymal transition
Claudin-1	↑	Enhanced epithelial integrity
Slug/Snail	↓	Suppressed invasion

Table 2. Antibody Validation Strategies

Parameter	Optimal Protocol
Specificity	siRNA knockdown + recombinant protein controls
Sensitivity	Titration series (0.5–2 µg/mL)
Reproducibility	Inter-lab validation using standardized lysates

Key Recommendations

Use UNC13C antibodies from suppliers providing epitope-mapping data (e.g., Abcam ab122725) .
For translational studies, prioritize antibodies validated in IHC and functional assays .
Address batch-to-batch variability by requesting lot-specific validation certificates .

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Unc13c Antibody