TRAIL/TNFSF10 Antibody [Unconjugated]

Western blot shows lysates of 786-O human renal cell adenocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TRAIL/TNFSF10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF375) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for TRAIL/TNFSF10 at approximately 27 and 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TRAIL/TNFSF10 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Goat Anti-Human TRAIL/TNFSF10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF375) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

TRAIL/TNFSF10 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human TRAIL/TNFSF10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF375) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

In the presence of a cross-linking antibody, Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050), and the metabolic inhibitor Actinomycin D, Recombinant Human TRAIL/TNFSF10 (Catalog # 375-TL) induces cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Under these conditions, cytotoxicity elicited by Recombinant Human TRAIL/TNFSF10 (12 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TRAIL/TNFSF10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF375). The ND50is typically <140 ng/mL.

IFN-beta -induced TRAIL expression in 2D-cultured cells. To determine the cells that express TRAIL and whose viability would be affected by IFN-beta , 2D-cultured HepG2 cells, primary liver sinusoidal endothelial cells (LSECs), LX-2 human hepatic stellate cells, and macrophages were treated with IFN-beta for 1 day. TRAIL expression was then analyzed via immunoblotting, and cell viability was determined using the WST-1 assay. (A) IFN-beta -induced TRAIL expression in the four cell types. The intensity of protein bands was quantified through densitometry using ImageJ (National Institutes of Health, //imagej.net/ij/index.html, 10 August 2023), and its relative expression was normalized to that of GAPDH. Data are presented as the mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01, and *** p < 0.001. (B) Cell viability upon IFN-beta treatment in the four cell types. Data are presented as the mean ± SD of three independent experiments. * p < 0.05 and ** p < 0.01. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/25/2/1325), licensed under a CC-BY license. Not internally tested by R&D Systems.

Suppression of IFN-beta -induced cell death by JAK inhibitors in liver tumor organoids. Liver tumor organoids were treated with JAK inhibitors (10 µM each of JAK1 and JAK2 inhibitor) for 20 min prior to IFN-beta treatment. After 24 h, the viability of organoids was analyzed using the WST-1 assay, and the expression of TRAIL and cleaved-caspase 3 (CC3) was analyzed by immunoblotting. (A) Viability of liver tumor organoids treated with IFN-beta and/or JAK inhibitors. (B) Expression of TRAIL and cleaved-caspase 3 (CC3) in liver tumor organoids treated with IFN-beta and/or JAK inhibitors. The results are presented as the mean ± standard deviation (SD) of three replicates. * p < 0.05 and *** p < 0.001; inh: inhibitor. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/25/2/1325), licensed under a CC-BY license. Not internally tested by R&D Systems.

Cell death in liver tumor organoids induced by ASC-IFN-beta . Adipose-derived stem cells (ASCs) were transiently transfected with pCMV3-IFN-beta . IFN-beta and TRAIL expression was determined via immunoblotting after 1 day. ASC-IFN-beta was added to liver tumor organoids on day 3, and ASC-IFN-beta -induced viability and cell death in liver tumor organoids were examined on day 4. (A) IFN-beta and TRAIL expression in ASC-IFN-beta . The intensity of protein expression was quantified through densitometry using ImageJ (National Institutes of Health, //imagej.net/ij/index.html, 10 August 2023), and the relative expression was normalized to that of GAPDH. (B) Suppression of liver tumor organoid viability by ASC-IFN-beta . The viability of organoids was analyzed using the WST-1 assay. (C) Cell death induced by ASC-IFN-beta in liver tumor organoids. ASC-IFN-beta -induced cell death was evaluated using a live (calcein-AM) and dead (ethidium homodimer-1) assay. In (C), the white boxes indicate areas that have been enlarged to distinguish the staining of the cells. White arrowheads indicate dead cells. Data shown are representative of three independent experiments. * p < 0.05 and ** p < 0.01. Scale bar: 500 µm. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/1422-0067/25/2/1325), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

TRAIL/TNFSF10 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human TRAIL/TNFSF10
  Thr95-Gly281
  Accession # P50591
• Specificity: Detects human TRAIL/TNFSF10 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse TRAIL is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TNFSF10
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization <140 ng/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for TRAIL/TNFSF10 Antibody [Unconjugated]

• Apo-2 ligand
• APO2L
• Apo-2Ltumor necrosis factor (ligand) family, member 10
• APO2Ltumor necrosis factor apoptosis-inducing ligand splice variant delta
• CD253 antigen
• CD253
• Protein TRAIL
• TL2
• TNF-related apoptosis-inducing ligand
• TNFSF10
• TRAIL
• TRAILTNF-related apoptosis inducing ligand TRAIL
• tumor necrosis factor (ligand) superfamily, member 10
• tumor necrosis factor ligand superfamily member 10

Background

TRAIL is a type II transmembrane protein belonging to the TNF superfamily and is now designated TNFSF10. TRAIL is active as a homotrimer and is produced by a variety of cell types in both the membrane bound form and also as a soluble molecule. It binds to any of the four TRAIL receptors as well as to Osteoprotegerin.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TRAIL/TNFSF10 Antibody (AF375)(17)

Publications using AF375

• Sánchez-Cerrillo, I;Agudo-Lera, M;Popova, O;Tsukalov, I;Calvet-Mirabent, M;de Los Santos, I;García-Fraile, L;Fuentes, P;Delgado-Arévalo, C;Alcain, J;Sánchez-Gaona, N;Grau-Expósito, J;Lázaro-Díez, M;Muñoz-Calleja, C;Alfranca, A;Genescà, M;Prado, JG;Vrbanac, V;Balazs, A;Buzón, MJ;Toribio, ML;Muñoz-Fernández, MA;Sánchez-Madrid, F;Martín-Gayo, E; Combined dendritic cell and anti-TIGIT immunotherapy potentiates adaptive NK cells against HIV-1 EMBO molecular medicine 2025-06-05 [PMID: 40473838] (Immunohistochemistry, Transgenic Mouse)
• Faust Akl, C;Andersen, BM;Li, Z;Giovannoni, F;Diebold, M;Sanmarco, LM;Kilian, M;Fehrenbacher, L;Pernin, F;Rone, JM;Lee, HG;Piester, G;Kenison, JE;Lee, JH;Illouz, T;Polonio, CM;Srun, L;Martinez, J;Chung, EN;Schüle, A;Plasencia, A;Li, L;Ferrara, K;Lewandrowski, M;Strathdee, CA;Lerner, L;Quéva, C;Clark, IC;Deneen, B;Lieberman, J;Sherr, DH;Antel, JP;Wheeler, MA;Ligon, KL;Chiocca, EA;Prinz, M;Reardon, DA;Quintana, FJ; Glioblastoma-instructed astrocytes suppress tumour-specific T cell immunity Nature 2025-05-21 [PMID: 40399681] (Immunohistochemistry, Human)
• Yoon, Y;Kim, CW;Kim, MY;Baik, SK;Jung, PY;Eom, YW; Interferon-? Overexpression in Adipose Tissue-Derived Stem Cells Induces HepG2 and Macrophage Cell Death in Liver Tumor Organoids via Induction of TNF-Related Apoptosis-Inducing Ligand Expression International journal of molecular sciences 2024-01-22 [PMID: 38279326] (Western Blot, Human)
• Luo, Y;Hao, H;Wang, Z;Ong, C;Dutcher, R;Xu, Y;Liu, J;Pedersen, LC;Xu, D; Heparan sulfate promotes TRAIL-induced tumor cell apoptosis bioRxiv : the preprint server for biology 2023-07-26 [PMID: 37546770] (Flow Cytometry, Human)
• Y An, J Jeon, L Sun, A Derakhshan, J Chen, S Carlson, H Cheng, C Silvin, X Yang, C Van Waes, Z Chen Death agonist antibody against TRAILR2/DR5/TNFRSF10B enhances birinapant anti-tumor activity in HPV-positive head and neck squamous cell carcinomas Scientific Reports, 2021-03-18;11(1):6392. 2021-03-18 [PMID: 33737574] (Neutralization, Human)
• LM Sanmarco, MA Wheeler, C Gutiérrez-, CM Polonio, M Linnerbaue, FA Pinho-Ribe, Z Li, F Giovannoni, KV Batterman, G Scalisi, SEJ Zandee, ES Heck, M Alsuwailm, DL Rosene, B Becher, IM Chiu, A Prat, FJ Quintana Gut-licensed IFN&amp;gamma+ NK cells drive LAMP1+TRAIL+ anti-inflammatory astrocytes Nature, 2021-01-06;0(0):. 2021-01-06 [PMID: 33408417] (IHC, Human)
• Chun Sung Byun, Soonjae Hwang, Sung-Hun Woo, Moon Young Kim, Jin Suk Lee, Jong In Lee et al. Adipose Tissue-Derived Mesenchymal Stem Cells Suppress Growth of Huh7 Hepatocellular Carcinoma Cells via Interferon (IFN)-beta -Mediated JAK/STAT1 Pathway in vitro International Journal of Medical Sciences 2020-02-18 [PMID: 32210710]
• T Li, Y Yang, H Song, H Li, A Cui, Y Liu, L Su, IN Crispe, Z Tu Activated NK cells kill hepatic stellate cells via p38/PI3K signaling in a TRAIL-involved degranulation manner J. Leukoc. Biol., 2019-02-12;0(0):. 2019-02-12 [PMID: 30748035] (Neutralization, Human)
• Mathieu J, Zhou W, Xing Y, Sperber H, Ferreccio A, Agoston Z, Kuppusamy K, Moon R, Ruohola-Baker H Hypoxia-inducible factors have distinct and stage-specific roles during reprogramming of human cells to pluripotency. Cell Stem Cell, 2014-03-20;14(5):592-605. 2014-03-20 [PMID: 24656769] (Neutralization, Human)
• Kim , Seong Mu, Woo , Ji Sun, Jeong , Chang Hy, Ryu , Chung He, Jang , Jae-Deog, Jeun , Sin-Soo Potential application of temozolomide in mesenchymal stem cell-based TRAIL gene therapy against malignant glioma. Stem Cells Transl Med, 2014-01-16;3(2):172-82. 2014-01-16 [PMID: 24436439] (Western Blot, Human)
• Siewiera J, El Costa H, Tabiasco J, Berrebi A, Cartron G, Le Bouteiller P, Jabrane-Ferrat N Human cytomegalovirus infection elicits new decidual natural killer cell effector functions. PLoS Pathog, 2013-04-04;9(4):e1003257. 2013-04-04 [PMID: 23592985] (Bioassay, Human)
• Navenot JM, Fujii N, Peiper SC KiSS1 metastasis suppressor gene product induces suppression of tyrosine kinase receptor signaling to Akt, tumor necrosis factor family ligand expression, and apoptosis. Mol. Pharmacol., 2009-02-06;75(5):1074-83. 2009-02-06 [PMID: 19201817] (Neutralization, Human)
• Rong S, Cai JH, Andrews J Cloning and apoptosis-inducing activities of canine and feline TRAIL. Mol. Cancer Ther., 2008-07-01;7(7):2181-91. 2008-07-01 [PMID: 18645027] (Affinity Purification, Western Blot, Canine, Feline, Human)
• Chamoux E, Houde N, L'Eriger K, Roux S Osteoprotegerin decreases human osteoclast apoptosis by inhibiting the TRAIL pathway. J. Cell. Physiol., 2008-08-01;216(2):536-42. 2008-08-01 [PMID: 18338379] (ICC, Neutralization, Human)
• Dunn C, Brunetto M, Reynolds G, Christophides T, Kennedy PT, Lampertico P, Das A, Lopes AR, Borrow P, Williams K, Humphreys E, Afford S, Adams DH, Bertoletti A, Maini MK Cytokines induced during chronic hepatitis B virus infection promote a pathway for NK cell-mediated liver damage. J. Exp. Med., 2007-03-12;204(3):667-80. 2007-03-12 [PMID: 17353365] (Neutralization, Human)
• Ryan LA, Peng H, Erichsen DA, Huang Y, Persidsky Y, Zhou Y, Gendelman HE, Zheng J TNF-related apoptosis-inducing ligand mediates human neuronal apoptosis: links to HIV-1-associated dementia. J. Neuroimmunol., 2004-03-01;148(1):127-39. 2004-03-01 [PMID: 14975593] (IHC-Fr, Human)
• Hueber A, Aduckathil S, Kociok N, Welsandt G, Dinslage S, Kirchhof B, Esser PJ Apoptosis-mediating receptor-ligand systems in human retinal pigment epithelial cells. Chin. Med. J., 2002-06-20;240(7):551-6. 2002-06-20 [PMID: 12136286] (IHC-Fr, Human)

Bioinformatics

• Gene Symbol: TNFSF10
• Uniprot:
  • P50591()