TNFRSF10C Antibody

What is TNFRSF10C and what is its biological function?

TNFRSF10C is a member of the TNF-receptor superfamily that contains an extracellular TRAIL-binding domain and a transmembrane domain, but lacks a cytoplasmic death domain . It is primarily localized to the cell membrane via GPI-anchor and lipid-anchor mechanisms . With a molecular weight of approximately 27kDa, TNFRSF10C plays a significant role in the regulation of programmed cell death and immune signaling pathways .

Unlike other TRAIL receptors capable of inducing apoptosis, TNFRSF10C functions as a decoy receptor that binds TRAIL ligand but does not transmit death signals intracellularly. This mechanism makes it a target of interest in cancer biology, as variations in its expression may influence cell survival and tumor progression .

What antibody formats are available for TNFRSF10C detection?

Several antibody formats have been developed for TNFRSF10C research:

• Monoclonal antibodies: Such as Human TRAIL R3/TNFRSF10C Antibody (Clone #90903, MAB630), which provides high specificity and reproducibility in applications like neutralization assays .

• Polyclonal antibodies: Including Goat Anti-Human TRAIL R3/TNFRSF10C Antigen Affinity-purified Polyclonal Antibody (AF630) and TNFRSF10C Rabbit Polyclonal Antibody (CAB1137), which offer broad epitope recognition .

• Recombinant fusion proteins: Recombinant Human TRAIL R3/TNFRSF10C Fc Chimera is utilized as a standard in functional assays and for antibody validation .

The choice between these formats depends on experimental requirements, with monoclonals preferred for reproducible detection of specific epitopes, while polyclonals provide stronger signals through multi-epitope binding.

What are the validated applications for TNFRSF10C antibodies?

TNFRSF10C antibodies have been validated for multiple research applications:

The efficacy of these applications depends on careful antibody validation and optimization for specific experimental conditions. Researchers should conduct preliminary titration experiments to determine optimal concentrations for their specific cell lines or tissue samples.

How should TNFRSF10C antibodies be optimized for Western blot applications?

For optimal Western blot detection of TNFRSF10C:

• Sample preparation: Use appropriate lysis buffers that preserve membrane proteins (such as RIPA buffer with protease inhibitors).

• Reducing conditions: TNFRSF10C detection has been successfully demonstrated under reducing conditions using Immunoblot Buffer Group 1 .

• Antibody concentration: Start with 0.5 μg/mL of anti-TNFRSF10C antibody (such as AF630) for PVDF membranes .

• Detection system: Use an appropriate HRP-conjugated secondary antibody, such as HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF019) for goat primary antibodies .

• Expected band size: The specific band for TNFRSF10C appears at approximately 27 kDa, correlating with its calculated molecular weight .

Troubleshooting tips include adjusting antibody concentration, extending incubation times, or using different blocking agents if non-specific binding occurs.

How can TNFRSF10C antibodies be utilized in TRAIL-mediated cytotoxicity studies?

TNFRSF10C antibodies are valuable tools for investigating TRAIL-mediated cytotoxicity pathways:

• Inhibition studies: Recombinant Human TRAIL R3/TNFRSF10C Fc Chimera inhibits Recombinant Human TRAIL/TNFSF10-induced cytotoxicity in L-929 mouse fibroblast cell lines in a dose-dependent manner .

• Neutralization assays: Under specific experimental conditions (1 μg/mL actinomycin D, 10-20 ng/mL Recombinant Human TRAIL R3/TNFRSF10C Fc Chimera, and 12-20 ng/mL Recombinant Human TRAIL/TNFSF10), anti-TNFRSF10C antibodies can neutralize the inhibitory effect of TRAIL R3 on TRAIL-induced cell death .

• Cross-linking studies: In certain experimental setups, additional cross-linking antibodies (such as Mouse polyHistidine Monoclonal Antibody at 5 μg/mL) may be required to enhance binding interactions .

The ND50 (neutralization dose) for antibody MAB630 is typically 0.02-0.08 μg/mL, while for AF630 it is 0.015-0.045 μg/mL, highlighting the potency of these research tools .

What is the significance of TNFRSF10C copy number variation in cancer research?

TNFRSF10C copy number variation (CNV) has emerged as a significant area of research in oncology:

• Metastatic association: TNFRSF10C CNV has been independently associated with distant metastatic disease in colorectal cancer (CRC). Multivariate analysis revealed that patients with TNFRSF10C CNV had significantly higher odds of metastatic disease [OR 4.63 (95% CI, 1.31–16.43); P=0.018] .

• Deletion patterns: The predominant CNV observed in TNFRSF10C is homozygous deletion, indicating copy number loss rather than amplification .

• Prognostic potential: In rectal cancer specifically, TNFRSF10C CNV showed even stronger association with metastatic disease [OR 5.69 (95% CI, 1.56–20.69); P=0.008], suggesting potential tissue-specific effects .

Researchers investigating TNFRSF10C in cancer should consider incorporating CNV analysis alongside protein expression studies using antibodies to gain a more comprehensive understanding of its role in disease progression.

What are the optimal storage and handling conditions for TNFRSF10C antibodies?

To maintain antibody functionality and extend shelf-life:

• Storage temperature: Store at -20 to -70°C for long-term storage (up to 12 months from receipt date) .

• After reconstitution:

  • For short-term use (up to 1 month): Store at 2-8°C under sterile conditions

  • For long-term storage (up to 6 months): Store at -20 to -70°C under sterile conditions

• Freeze-thaw cycles: Use manual defrost freezers and avoid repeated freeze-thaw cycles that can degrade antibody quality .

• Reconstitution protocols: Follow manufacturer's specific instructions for reconstitution buffer composition and concentration.

• Working aliquots: Upon receipt, consider preparing small working aliquots to minimize freeze-thaw cycles for the stock solution.

How can cross-reactivity issues be addressed when using TNFRSF10C antibodies?

Cross-reactivity is a common challenge in antibody-based research:

• Species specificity: Different antibodies show varied species reactivity. For example, the CAB1137 antibody demonstrates reactivity with mouse and rat samples , while others are human-specific.

• Validation approaches:

  • Positive controls: Use known positive samples such as mouse liver or rat liver for cross-reactivity testing .

  • Blocking peptides: Consider using immunizing peptides to confirm specificity.

  • Knockout/knockdown validation: When available, cells with TNFRSF10C knockout or knockdown provide definitive specificity controls.

• Background reduction: Optimize blocking conditions (5% BSA or milk) and include appropriate detergents in wash buffers to minimize non-specific binding.

• Antibody dilution: Start with manufacturer-recommended dilutions (e.g., 1:500 - 1:1000 for Western blot with CAB1137 ) and adjust as needed based on signal-to-noise ratio.

How can TNFRSF10C antibodies be used to investigate apoptosis resistance in cancer models?

TNFRSF10C, as a decoy receptor for TRAIL, has implications for cancer cell survival:

• Expression correlation studies: Use antibodies in IHC or Western blot to correlate TNFRSF10C expression levels with apoptosis resistance in different cancer cell lines or patient samples.

• Mechanistic investigations: Combine antibody detection with functional assays to determine whether TNFRSF10C overexpression correlates with resistance to TRAIL-induced apoptosis.

• Therapeutic targeting: Neutralizing antibodies against TNFRSF10C could potentially sensitize resistant cancer cells to TRAIL therapy by blocking the decoy receptor function.

• Biomarker development: TNFRSF10C detection might serve as a predictive biomarker for TRAIL therapy response, with expression levels determined using validated antibodies.

• Multi-parameter analysis: Combine TNFRSF10C detection with analysis of other TRAIL pathway components to develop comprehensive profiles of apoptosis sensitivity.

What is the role of TNFRSF10C in metastatic progression and how can antibodies help elucidate this?

Based on CNV studies showing association with metastasis:

• Expression analysis: Anti-TNFRSF10C antibodies can be used to examine protein expression patterns in primary tumors versus metastatic lesions through techniques like tissue microarray analysis.

• Mechanistic studies: Researchers can investigate whether TNFRSF10C affects metastatic potential through:

  • Cell migration/invasion assays after modulating TNFRSF10C expression

  • Correlation of TNFRSF10C expression with epithelial-mesenchymal transition markers

  • Analysis of TNFRSF10C expression in circulating tumor cells

• Clinical correlation: Combined genomic (CNV) and proteomic (antibody-based) analyses can provide more comprehensive insights into TNFRSF10C's role in metastasis.

• Prognostic value: The strong association between TNFRSF10C CNV and metastatic disease [OR 4.63 (95% CI, 1.31–16.43)] suggests potential as a prognostic marker, which can be further validated using antibody-based approaches .