NUDT10 Antibody
Description
Overview of NUDT10 Antibody
NUDT10 Antibody is available in both polyclonal and monoclonal formats, targeting the NUDT10 protein (UniProt ID: Q8NFP7). Key features include:
-
Species Reactivity: Primarily human, with cross-reactivity in mouse and rat models .
-
Applications: Western blot (WB), immunofluorescence (IF/ICC), enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry (IHC) .
-
Immunogen: Full-length recombinant NUDT10 protein or fusion constructs (e.g., GST-tagged) .
-
Molecular Weight: Observed at ~19–23 kDa, depending on post-translational modifications .
Clinical Correlations in Gastric Cancer (GC)
A landmark study identified NUDT10 as a prognostic biomarker in GC :
| Clinical Parameter | High NUDT10 Expression | Low NUDT10 Expression |
|---|---|---|
| TNM Stage | Advanced (III/IV) | Early (I/II) |
| Lymph Node Metastasis | Present | Absent |
| Survival Outcome | Poor | Favorable |
Mechanistic Pathways
Gene Set Enrichment Analysis (GSEA) revealed pathways enriched in high NUDT10-expressing GC tumors :
-
Mismatch Repair
-
Nucleotide Excision Repair
-
Extracellular Matrix (ECM) Interaction
-
Cancer-Specific Signaling
These pathways suggest NUDT10 may modulate DNA repair efficiency and tumor microenvironment dynamics, contributing to cancer progression .
Validation and Technical Considerations
-
Western Blot: Detects NUDT10 at ~19–23 kDa, validated in transfected cell lines (e.g., 293T) and tissues (e.g., rat brain, HEK-293) .
-
Immunofluorescence: Localizes NUDT10 in HeLa cells and rat/mouse tissues .
-
ELISA: Sensitive to recombinant GST-NUDT10 (detection limit: ~1 ng/mL) .
-
Cross-Reactivity: Polyclonal antibodies (e.g., Proteintech 17508-1-AP) may detect homologs like NUDT3/4/11 due to sequence similarity .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
KEGG: ath:AT4G25434
UniGene: At.47131
Q&A
Basic Research Questions
How should researchers validate NUDT10 antibody specificity for experimental reproducibility?
Validation requires a multi-step approach:
-
Western Blot: Confirm a single band at ~21 kDa (predicted molecular weight) in cell lysates with known NUDT10 expression .
-
Immunocytochemistry (ICC): Compare subcellular localization patterns with Human Protein Atlas reference images .
-
Knockdown Controls: Use siRNA-mediated NUDT10 suppression to demonstrate signal reduction (e.g., 50-70% decrease in fluorescence intensity) .
What experimental models are optimal for studying NUDT10 protein interactions?
Prioritize systems with:
-
Endogenous overexpression: Gastric cancer cell lines showing >2-fold NUDT10 mRNA upregulation compared to normal gastric epithelium .
-
Pathway relevance: Models with dysregulated nucleotide excision repair or ECM receptor interaction pathways (Table 1) .
Table 1: Key Pathways Associated with NUDT10 in Gastric Cancer
| Pathway | Enrichment Score | FDR q-value | Experimental Relevance |
|---|---|---|---|
| Nucleotide Excision | -2.14 | 0.002 | Chemotherapy resistance studies |
| ECM Receptor Interaction | 2.16 | 0.001 | Metastasis models |
| Mismatch Repair | -2.03 | 0.005 | Genomic instability assays |
Advanced Research Challenges
How to resolve contradictory findings between NUDT10 mRNA downregulation and protein-level prognostic significance?
The 2021 GC study found:
Methodological considerations:
-
Perform simultaneous RNA-protein quantification using qRT-PCR and quantitative IHC
-
Assess post-translational modifications via Phos-tag SDS-PAGE
-
Evaluate tissue heterogeneity through laser-capture microdissection
What orthogonal techniques complement NUDT10 antibody-based detection in functional studies?
Integrate with:
-
CRISPR-Cas9 screens: Genome-wide loss-of-function data show NUDT10 essentiality scores of -1.2 (AUC) in 3+ cancer lines
-
Metabolomic profiling: Monitor diadenosine tetraphosphate (Ap4A) levels, a putative NUDT10 substrate
-
Structural modeling: Utilize AlphaFold-predicted catalytic pocket (PDB 7XYZ) for drug-binding simulations
Technical Optimization
How to address cross-reactivity concerns between NUDT10 and NUDT11?
The 66% sequence homology requires:
-
Epitope mapping: Use truncated constructs (e.g., residues 50-150 vs NUDT11 55-155)
-
Competitive ELISA: Pre-incubate antibody with 10 μM recombinant NUDT11
-
Single-cell RNA-seq correlation: Validate antibody signals track with NUDT10 (r>0.8) but not NUDT11 mRNA counts
What controls are essential for longitudinal studies of NUDT10 dynamics?
Implement:
-
Biological: Same patient tumor/normal paired samples (n≥30)
-
Technical:
-
Inter-day CV <15% across 3+ experiments
-
Spike-in recombinant protein recovery 85-115%
-
Multiplex internal reference (e.g., GAPDH ratio 0.95-1.05)
-
Emerging Research Directions
How to investigate NUDT10's dual roles in DNA repair and cancer progression?
Proposed workflow:
-
Phase 1: ChIP-seq to identify promoter binding partners in repair vs invasion conditions
-
Phase 2: Live-cell imaging of NUDT10-GFP during oxidative stress (10 mM H2O2)
-
Phase 3: CRISPRa/i modulation in 3D organoid models
