TNFRSF10D Antibody

What is TNFRSF10D and what is its biological role?

TNFRSF10D (Tumor Necrosis Factor Receptor Superfamily Member 10D), also known as Decoy Receptor 2 (DcR2) or TRAIL Receptor 4 (TRAIL-R4), is a type I transmembrane protein and member of the TNF receptor family. It functions as a receptor for TRAIL (TNF-Related Apoptosis-Inducing Ligand/APO2L). TNFRSF10D is unique among TRAIL receptors as it contains a truncated death domain in its cytoplasmic portion, making it incapable of inducing apoptosis . Instead, it serves as a decoy receptor that can protect cells from TRAIL-mediated apoptosis by competing with functional death receptors for TRAIL binding. When overexpressed, TNFRSF10D can protect cells bearing TRAIL R1 and/or TRAIL R2 from TRAIL-induced apoptotic signaling .

Where is TNFRSF10D predominantly expressed in human tissues?

TNFRSF10D is widely expressed throughout human tissues with particularly high expression in fetal kidney, lung, and liver, as well as adult testis and liver. It is also expressed in peripheral blood leukocytes, colon, small intestine, ovary, prostate, thymus, spleen, pancreas, kidney, lung, placenta, and heart . Flow cytometry data shows that TNFRSF10D is detectable on human blood granulocytes, where it can be visualized using specific monoclonal antibodies .

How does TNFRSF10D differ from other TRAIL receptors?

TNFRSF10D differs from other TRAIL receptors (TNFRSF10A/DR4 and TNFRSF10B/DR5) primarily in its cytoplasmic domain structure. While functional death receptors contain intact death domains capable of initiating apoptotic signaling cascades, TNFRSF10D contains a truncated death domain motif . This structural difference prevents TNFRSF10D from inducing programmed cell death upon ligand binding. Additionally, there are contradictory reports regarding TNFRSF10D's ability to activate the NF-kappa-B pathway, with some studies suggesting it cannot while others indicate it can induce this signaling pathway .

What are the most validated applications for TNFRSF10D antibodies in research?

Based on the extensive validation data from multiple suppliers, TNFRSF10D antibodies have been successfully employed in several experimental applications:

The optimal dilution ranges differ by application and antibody source but typically include: 1:500-1:1000 for WB, 1:50-1:200 for IHC-P, and 1:10-1:50 for flow cytometry .

What controls should be included when validating a new TNFRSF10D antibody?

When validating a new TNFRSF10D antibody, several controls should be included to ensure specificity and reliability:

• Isotype controls: For example, using Mouse IgG1 isotype control antibody (e.g., Catalog # IC002A) when working with mouse monoclonal anti-TNFRSF10D antibodies .

• Positive tissue/cell controls: Human peripheral blood granulocytes serve as excellent positive controls for flow cytometry applications as they express detectable levels of TNFRSF10D .

• Recombinant protein controls: Recombinant Human TRAILR4/TNFRSF10D Fc Chimera protein can be used as a standard for ELISA development and as a positive control in Western blot applications .

• Cross-reactivity assessment: Test against other TRAIL receptors to confirm specificity. For example, ensure less than 5% cross-reactivity with recombinant human TRAIL R1 in sandwich immunoassays .

• Blocking peptides: When available, using the immunogenic peptide to pre-absorb the antibody can confirm signal specificity.

How can I optimize detection of TNFRSF10D in flow cytometry experiments?

To optimize detection of TNFRSF10D in flow cytometry experiments:

• Cell preparation: For peripheral blood samples, isolate granulocytes using standard density gradient separation methods. Freshly isolated cells typically provide better results than frozen samples .

• Antibody titration: Perform careful titration experiments to determine optimal antibody concentration. Starting dilutions of 1:10-1:50 are recommended for most flow cytometry applications of TNFRSF10D antibodies .

• Multi-color panel design: Consider using TNFRSF10D antibodies in conjunction with lineage markers. For example, CEACAM-8/CD66b can be used alongside TNFRSF10D detection to better characterize granulocyte populations .

• Protocol optimization: Follow standardized protocols for staining membrane-associated proteins, such as those provided by antibody manufacturers. Gentle fixation and permeabilization methods are recommended to preserve epitope integrity .

• Controls: Always include appropriate isotype controls (matching the isotype of your TNFRSF10D antibody) and unstained controls to accurately set gates and differentiate positive from negative populations .

How can TNFRSF10D antibodies be used to study TRAIL resistance mechanisms in cancer?

TNFRSF10D antibodies have proven valuable in elucidating TRAIL resistance mechanisms in cancer:

• Expression analysis: TNFRSF10D is often overexpressed in cancer cells resistant to TRAIL-induced apoptosis. Flow cytometry with TNFRSF10D antibodies can quantify expression levels and correlate them with resistance profiles .

• Mechanism investigation: Studies have shown that TNFRSF10D can protect cells bearing TRAIL R1 and/or TRAIL R2 from TRAIL-mediated apoptosis. Neutralizing antibodies against TNFRSF10D can reverse this protection in functional assays, helping to determine the contribution of this receptor to resistance mechanisms .

• Therapeutic targeting: TNFRSF10D antibodies can be used to assess potential therapeutic strategies targeting this decoy receptor. For example, the study cited in search result explored how oxaliplatin resistance in colorectal cancer enhances TRAIL sensitivity via death receptor 4 upregulation and lipid raft localization.

• Combination therapy assessment: Researchers can use these antibodies to investigate how cancer treatments affect TNFRSF10D expression and function. For instance, studies have shown that alternol sensitizes renal carcinoma cells to TRAIL-induced apoptosis, and TNFRSF10D antibodies help elucidate these mechanisms .

What is the role of TNFRSF10D in immune cell function and how can antibodies help investigate this?

TNFRSF10D plays important roles in immune cell function that can be investigated using antibodies:

• Dendritic cell interactions: TNFRSF10D antibodies have helped identify that plasmacytoid dendritic cells express TRAIL and induce CD4+ T-cell apoptosis in HIV-1 viremic patients . Flow cytometry with these antibodies can characterize the expression patterns on different immune cell populations.

• Immune regulation: TNFRSF10D may contribute to immune homeostasis by preventing excessive TRAIL-mediated apoptosis in immune cells. Neutralizing antibodies can help determine the functional significance of this protection .

• Cytokine responses: Studies using TNFRSF10D antibodies have shown that TRAIL+ human plasmacytoid dendritic cells kill tumor cells in vitro, and this mechanism is influenced by imiquimod and IFN-alpha. The antibodies allow for correlation between receptor expression and functional outcomes .

• Myeloid cell development: Research has linked TNFRSF family members to myeloid cell development and maturation. TNFRSF10D antibodies can help track expression changes during differentiation and activation of these cells .

How do pH changes affect TNFRSF10D expression and function in cancer cells?

Recent research cited in the search results indicates that extracellular pH influences TRAIL-induced signaling in cancer cells, with potential implications for TNFRSF10D:

• Expression regulation: Studies investigating TRAIL-induced signaling in pancreatic ductal adenocarcinoma cells have shown that extracellular pH can affect the expression levels of TRAIL receptors, including TNFRSF10D .

• Functional impact: Changes in pH can alter the sensitivity of cancer cells to TRAIL-mediated apoptosis, potentially by affecting the distribution or functionality of decoy receptors like TNFRSF10D .

• Experimental approach: To study this phenomenon, researchers can use TNFRSF10D antibodies in flow cytometry or Western blot analyses to quantify expression levels under different pH conditions. Additionally, neutralizing antibodies can help determine if the pH-dependent effects are mediated through TNFRSF10D or other TRAIL receptors .

• Clinical relevance: Understanding how tumor microenvironment acidity affects TRAIL receptor expression may inform therapeutic strategies targeting these pathways in cancer treatment .

What are the key storage and handling requirements for TNFRSF10D antibodies?

Proper storage and handling of TNFRSF10D antibodies is crucial for maintaining their functionality:

• Storage temperature: Most TNFRSF10D antibodies should be stored at -20°C for up to 1 year from the date of receipt. Some conjugated antibodies should be stored at 2-8°C and should not be frozen .

• Avoid freeze-thaw cycles: Repeated freeze-thaw cycles can significantly reduce antibody activity. It is recommended to aliquot antibodies before freezing to minimize the number of freeze-thaw cycles .

• Light protection: For fluorophore-conjugated antibodies (e.g., APC or PE-conjugated anti-TNFRSF10D), protection from light is essential to prevent photobleaching .

• Reconstitution: Lyophilized antibodies should be reconstituted according to manufacturer specifications, typically in sterile water or buffer solutions. After reconstitution, they can be stored at 2-8°C for 1 month or aliquoted and stored at -20 to -70°C for 6 months under sterile conditions .

• Preservatives: Many antibody formulations contain 0.02-0.09% sodium azide as a preservative. This should be considered when designing experiments where enzyme activity might be affected by azide .

How can I troubleshoot weak or non-specific staining in flow cytometry with TNFRSF10D antibodies?

When encountering weak or non-specific staining in flow cytometry with TNFRSF10D antibodies, consider these troubleshooting approaches:

• Antibody titration: Suboptimal antibody concentration can lead to weak or non-specific signals. Perform a titration experiment to determine the optimal concentration for your specific application .

• Cell viability: Ensure high cell viability before staining, as dead or dying cells can contribute to background staining. Include a viability dye in your panel to exclude dead cells from analysis .

• Fc receptor blocking: Non-specific binding to Fc receptors on immune cells can occur. Use appropriate Fc receptor blocking reagents before adding the TNFRSF10D antibody .

• Buffer optimization: The staining buffer composition can affect antibody binding. Try different buffers with varying BSA or serum concentrations to reduce background .

• Compensation controls: For multi-color flow cytometry, proper compensation is essential. Use single-stained controls for each fluorochrome in your panel to set up accurate compensation matrices .

• Fixation effects: Some epitopes may be sensitive to certain fixation protocols. If staining is weak after fixation, try alternative fixation methods or perform staining on live cells if possible .

What considerations should be made when selecting between polyclonal and monoclonal TNFRSF10D antibodies?

The choice between polyclonal and monoclonal TNFRSF10D antibodies depends on the specific research application:

For critical experiments, it is advisable to validate findings using both monoclonal and polyclonal antibodies to ensure robust results .

How does TNFRSF10D expression correlate with cancer progression and therapeutic response?

Research using TNFRSF10D antibodies has revealed important correlations between receptor expression and cancer behavior:

• Therapeutic resistance: Studies have shown that TNFRSF10D expression can protect cancer cells from TRAIL-induced apoptosis, potentially contributing to treatment resistance. Flow cytometry with anti-TNFRSF10D antibodies can quantify expression levels in patient samples and correlate them with clinical outcomes .

• Cancer stem cells: Research has demonstrated that CD133+ cancer stem cells in non-small cell lung cancer can be targeted by mesenchymal stem cells expressing TRAIL, with TNFRSF10D potentially playing a role in modulating this sensitivity .

• p53 status interaction: The impact of p53 status on TRAIL-mediated apoptotic and non-apoptotic signaling in cancer cells has been investigated using TNFRSF10D antibodies, revealing complex interactions between tumor suppressor pathways and TRAIL receptor signaling .

• Chemotherapy synergy: Studies employing TNFRSF10D antibodies have shown that cisplatin sensitizes human hepatocellular carcinoma cells, but not hepatocytes and mesenchymal stem cells, to TRAIL within a therapeutic window partially depending on the upregulation of death receptors and potentially downregulation of decoy receptors like TNFRSF10D .

What is the significance of TNFRSF10D in immune-related disorders?

TNFRSF10D plays important roles in immune regulation that may contribute to various disorders:

• HIV pathogenesis: Research using TNFRSF10D antibodies has identified that plasmacytoid dendritic cells express TRAIL and induce CD4+ T-cell apoptosis in HIV-1 viremic patients, potentially contributing to T-cell depletion .

• Inflammatory conditions: As a member of the TNFRSF family, TNFRSF10D may contribute to inflammatory processes by modulating TRAIL-mediated signaling. Antibodies against TNFRSF10D can help elucidate these mechanisms in various inflammatory disease models .

• Autoimmunity: Studies in animal models have shown that TRAIL deficiency is associated with accelerated autoimmune diseases, suggesting that the balance between functional death receptors and decoy receptors like TNFRSF10D may influence autoimmune processes .

• Immune cell development: Research using knockout mouse models of various TNFRSF members has revealed roles in immune cell development and function. While specific TNFRSF10D knockout phenotypes are not reported in the provided materials, antibodies against this receptor can help map its expression patterns during immune cell differentiation and activation .

How can TNFRSF10D antibodies be used to develop new therapeutic strategies?

TNFRSF10D antibodies play important roles in developing new therapeutic approaches:

• Target validation: Antibodies help validate TNFRSF10D as a potential therapeutic target by confirming its expression in disease states and elucidating its functional role .

• Neutralization strategies: Neutralizing antibodies against TNFRSF10D can reverse its protective effect against TRAIL-induced apoptosis, potentially sensitizing cancer cells to TRAIL-based therapies. In vitro assays measuring this effect use antibodies like the goat anti-human TRAIL R4/TNFRSF10D antibody described in search result .

• Companion diagnostics: TNFRSF10D antibodies could be developed into diagnostic assays to identify patients most likely to benefit from TRAIL receptor-targeting therapies based on their decoy receptor expression profiles .

• Antibody-drug conjugates: While not explicitly mentioned in the search results, the specific binding of TNFRSF10D antibodies to cancer cells that overexpress this receptor could potentially be leveraged for antibody-drug conjugate development .

• Immune modulation: Understanding how TNFRSF10D impacts immune cell function using antibody-based detection methods could inform strategies to modulate immune responses in cancer, autoimmunity, and infectious diseases .