tbx-30 Antibody

Here’s a structured, research-oriented FAQ collection for TBX3 antibody applications in academic contexts, synthesized from peer-reviewed studies and technical resources:

Advanced Research Questions

• How to resolve contradictory data on TBX3’s role in Wnt/β-catenin signaling?

  • Context-dependent effects require:

    Model System	TBX3 Activity	Key Readout	Citation
    HEK293T (Wnt-OFF)	Represses STF reporter	↓ Luciferase
    HCT116 (β-catenin mutant)	Enhances proliferation	↑ Metastasis in zebrafish
    Melanoma (TBX3-high)	Promotes invasiveness	↓ E-cadherin

  • Design dose-response assays with pathway inhibitors (e.g., XAV939 for β-catenin degradation) .

• What methodologies detect TBX3’s interaction with chromatin in stem cells?

  • Combine ChIP-seq (anti-TBX3 antibody AF0319 ) with CUT&RUN for high-resolution binding site mapping .

  • Validate targets (e.g., Zscan4, Tcstv1) via:

    • Dual-luciferase reporters with TBX3-binding site mutations

    • CRISPRa/i perturbation followed by RNA-seq

Technical Challenges & Solutions

• Interpreting multiple bands in TBX3 Western blots

  • Expected isoforms:

    Band (kDa)	Isoform	Functional Relevance
    75	TBX3-α	Full-length transcriptional repressor
    68	TBX3-β	Truncated, dominant-negative form

  • Use phosphatase inhibitors (e.g., PhosSTOP) to reduce phosphorylation-induced mobility shifts .

• Selecting in vivo models for TBX3 functional studies

  • Zebrafish xenografts: Quantify metastatic spread of TBX3-OE CRC cells (HCT116) via fluorescence microscopy .

  • THX humanized mice: Assess antibody-dependent cellular toxicity using TBX3+ lymphoma cells (Daudi line) .

Data Interpretation Framework

• Discrepancies in TBX3’s role in senescence escape

  • Mechanistic divergence:

    	Breast Cancer	Melanoma
    Target	ARF repression	p21 bypass
    Assay	SA-β-gal	EdU incorporation

  • Standardize senescence markers (p16INK4a, Lamin B1) across models.