TRAILR2/TNFRSF10B Antibody

Western Blot: TRAILR2/TNFRSF10B Antibody [NB100-56618] - TRAIL R2/TNFRSF10B Antibody [NB100-56618] - Total protein from HL-60, HepG2, HCT-116 and human placenta was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-TRAIL R2 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunohistochemistry-Paraffin: TRAILR2/TNFRSF10B Antibody [NB100-56618] - TRAIL R2/TNFRSF10B Antibody [NB100-56618] - Analysis of a formalin fixed paraffin-embedded (FFPE) human brain cerebellum using 1:200 conc. of TRAIL R2/TNFRSF10B antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 30 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 9.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 15 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Cytoplasmic staining was observed in the Purkinje cell layer.

Immunohistochemistry: TRAILR2/TNFRSF10B Antibody [NB100-56618] - Pharmacodynamic assays for ONC201. IHC analyses of CHOP, TRAILR2/TNFRSF10B, and double-stranded DNA breaks (TUNEL) in baseline and ONC201-treated biopsies for an endometrial cancer patient. The biopsy was done 9.8 months after starting ONC201 treatment (7 days after the most recent ONC201 dose). Image collected and cropped by CiteAb from the following publication (//jitc.biomedcentral.com/articles/10.1186/s40425-019-0599-8) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: TRAIL R2/TNFRSF10B Antibody [NB100-56618] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-TRAIL R2 (H76) at 5ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: TRAILR2/TNFRSF10B Antibody [NB100-56618] - TRAIL R2/TNFRSF10B Antibody [NB100-56618] - Analysis using the Azide Free version of NB100-56618. Detection of 20 ug of whole cell lysates from HL60 cells with anti-TRAIL-R2 at 5 ug/ml.

• Reactivity: Hu, Mu
• Applications: WB, Flow, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

TRAILR2/TNFRSF10B Antibody Summary

• Immunogen: Rabbit anti-DR5 polyclonal antibody was raised against a peptide corresponding to amino acids 388 to 407 (isoform 2) of human DR5 precursor (1,2). The same sequence is found in isoform 1 at amino acids 417-436.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TNFRSF10B
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Flow (Cell Surface) reported in scientific literature (KOYAMA (2003); PMID 16426839)
  • Flow Cytometry 1:20-1:2000
  • Immunocytochemistry/Immunofluorescence 1:10-1:500. Use reported in scientific literature (Georgakis et al (2005))
  • Immunohistochemistry 1:200. Use reported in scientific literature (PMID 27740879)
  • Immunohistochemistry-Paraffin reported in scientific literature (Younes et al (2006))
  • Western Blot 5 ug/ml
• Control:

  Jurkat Whole Cell Lysate
  TRAILR2/TNFRSF10B Overexpression Lysate

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 21474767).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for TRAILR2/TNFRSF10B Antibody

• CD262 antigen
• CD262
• death domain containing receptor for TRAIL/Apo-2L
• Death receptor 5
• DR5
• DR5TRICK2B
• Fas-like protein
• KILLER/DR5
• KILLERapoptosis inducing protein TRICK2A/2B
• TNF-related apoptosis-inducing ligand receptor 2
• TNFRSF10B
• TRAIL R2
• TRAIL receptor 2
• TRAILR2
• TRAIL-R2apoptosis inducing receptor TRAIL-R2
• TRAILR2cytotoxic TRAIL receptor-2
• TRICK2
• TRICK2A
• TRICKB
• tumor necrosis factor receptor superfamily member 10B
• tumor necrosis factor receptor superfamily, member 10b
• tumor necrosis factor receptor-like protein ZTNFR9
• ZTNFR9

Background

Apoptosis is induced by certain cytokines including TNF and Fas ligand in the TNF family through their death domain containing receptors. TRAIL/Apo2L is a new member of the TNF family. DR4 was recently identified as the receptor for TRAIL. A novel death domain containing receptor for TRAIL was more recently identified and designated DR5, Apo2, TRAIL-R2, TRICK2, or KILLER by several groups independently (1-2). Like DR4, DR5 transcript is widely expressed in normal tissues and in many types of tumor cells. DR5 binds to TRAIL and mediates TRAIL induced cell death. Overexpression of DR5 induces apoptosis and activates NF-kB.

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 TRAILR2/TNFRSF10B Antibody (NB100-56618)(41)

Publications using NB100-56618

• Chen L, Hao M, Yan J et al. Citrus-derived DHCP inhibits mitochondrial complex II to enhance TRAIL sensitivity via ROS-induced DR5 upregulation The Journal of biological chemistry 2021-03-04 [PMID: 33676890] (FLOW, Human)
• Arrillaga-Romany I, Odia Y et al. Biological activity of weekly ONC201 in adult recurrent glioblastoma patients. Neuro Oncol 2020-11-01 [PMID: 31702782] (IF/IHC, Human)
• Stein M N, Malhotra J et al. Safety and enhanced immunostimulatory activity of the DRD2 antagonist ONC201 in advanced solid tumor patients with weekly oral administration. J Immunother Cancer 2019-05-22 [PMID: 31118108] (IF/IHC, Human)
• Xue M, Ge Y et al Apoptosis is induced by docosahexaenoic acid in breast cancer cells via death receptor and mitochondria-mediated pathways Mol Med Rep 2017-06-07 [PMID: 28586001] (WB, Human)
  
  Details:
  Citation using the Azide Free version of this antibody.
• Shin GC, Kang HS, Lee AR, Kim KH Hepatitis B virus-triggered autophagy targets TNFRSF10B/death receptor 5 for degradation to limit TNFSF10/TRAIL response. Autophagy 2016-10-14 [PMID: 27740879] (WB, IF/IHC, Human)
• Allen JE, Crowder R, El-Deiry WS. First-In-Class Small Molecule ONC201 Induces DR5 and Cell Death in Tumor but Not Normal Cells to Provide a Wide Therapeutic Index as an Anti-Cancer Agent. PLoS ONE. 2015-11-20 [PMID: 26580220] (FLOW, Human)
  
  Details:
  DR5 (TRAIL R2/TNFRSF10B) antibody was used for FLOW analysis of HCT116 (tumor) and HFF (normal) cells and the protocol implicated - fixation of cells in 4% paraformaldehyde-PBS for 20 minutes, use of primary at 1:250 dilution with ON incubation, detection using Alexa fluor 488 conjugated anti-rabbit secondary (Fig 2).
• Liu Y, Hawkins OE, Vilgelm AE et al. Combining an Aurora Kinase Inhibitor and a Death Receptor Ligand/Agonist Antibody Triggers Apoptosis in Melanoma Cells and Prevents Tumor Growth in Preclinical Mouse Models Clin. Cancer Res. 2015-07-07 [PMID: 26152738] (WB, Human)
  
  Details:
  TRAIL R2/TNFRSF10B (DR5) antibody was used for WB on lysates of human melanoma cell line Hs294T treated with mock siRNA or DR5-siRNAs (Figure 3D).
• Foster KA, Jane EP, Premkumar DR et al. NVP-BKM120 potentiates apoptosis in tumor necrosis factor-related apoptosis-inducing ligand-resistant glioma cell lines via upregulation of Noxa and death receptor 5 Int. J. Oncol. 2015-06-05 [PMID: 26044191] (ICC/IF, Human)
• Talekar MK, Allen JE, Dicker DT, El-Deiry WS. ONC201 induces cell death in pediatric non-Hodgkin's lymphoma cells Cell Cycle. 2015-06-01 [PMID: 26030065] (FLOW)
  
  Details:
  DR5 antibody (Imgenex Img-120A) was used at 1:200 dilution for FLOW applictaion in experiments involving ONC201 induced cell death in cell lines namely Granta, UPN2, NCEB, Karpas299, BJAB, Daudi cell lines (Fig. S1A, B).
• Prabhu VV, Allen JE, Dicker DT, El-Deiry WS. Small molecule ONC201/TIC10 targets chemotherapy-resistant colorectal cancer stem-like cells in an Akt/Foxo3a/TRAIL-dependent manner Cancer Res. 2015-02-20 [PMID: 25712124] (FLOW, Human)
  
  Details:
  TRAIL R2/TNFRSF10B antibody was used for WB and FLOW applications in experiments involving colon cancer cell lines namely SW480, DLD1, and HCT116 cells and HCT116 cells overexpressing myristoylated Akt. For FLOW, the cells were fixed in 4% paraformaldehyde, incubated with primary antibody at 1:100 dilution or with normal IgG control for ON or 2 hours at 4C, washed and incubated with secondary antibody which followed analysis on Elite ESP flow cytometer (Supplementary Figure S5). WB performed on cancer cells subjected or not to SiRNA knockdown of DR5 (Supplementary Figure S6 ).
• Xu J, Zhou JY, Xu Z et al. The role of Cullin3-mediated ubiquitination of the catalytic subunit of PP2A in TRAIL signaling Cell Cycle. 2015-01-01 [PMID: 25551360] (WB, Human)
  
  Details:
  TRAIL R2/TNFRSF10B (DR5) antibody used for WB on DISC-Flag immunoprecipitates from lysates of SUM159 cells (human breast cell line) that were treated with 100 ng/ml Flag-TRAIL, 1 mg/ml Enhancer and 2 mg/ml anti-Flag (Flag/M2) for 30 min at 37 C (Figure 3C).
• Chen Jun-Jie, Bozza William P, Di Xu et al. H-Ras regulation of TRAIL death receptor mediated apoptosis. Oncotarget. 2014-07-15 [PMID: 25026275] (Human)
• Qiu F, Hu M, Tang B, Liu X et al. Annexin V-TRAIL fusion protein is a more sensitive and potent apoptotic inducer for cancer therapy. Sci Rep 2013-12-20 [PMID: 24356445] (WB, Human)
• Xu J, Xu Z, Zhou JY et al. Regulation of the Src-PP2A Interaction in Tumor Necrosis Factor (TNF)-related Apoptosis-inducing Ligand (TRAIL)-induced Apoptosis. J Biol Chem 2013-11-15 [PMID: 24100030] (WB, Human)
  
  Details:
  BT549 human breast carcinoma cells (lipid raft and non-raft membrane fractions), Fig 5A, 5B;DR5 was more highly detected in Non-raft than raft membrane fractions.
• Abdulghani Junaid, Allen Joshua E, Dicker David T et al. Sorafenib sensitizes solid tumors to Apo2L/TRAIL and Apo2L/TRAIL receptor agonist antibodies by the Jak2-Stat3-Mcl1 axis. PLoS One. 2013-01-01 [PMID: 24086526] (FLOW, Human)
• Di X, Zhang G, Zhang Y et al. Accumulation of autophagosomes in breast cancer cells induces TRAIL resistance through downregulation of surface expression of death receptors 4 and 5. Oncotarget 2013-09-01 [PMID: 23988408] (ICC/IF)
• Allen JE, Ferrini R, Dicker DT et al. Targeting TRAIL death receptor 4 with trivalent DR4 Atrimer complexes. Mol Cancer Ther. 2012-10-01 [PMID: 22802267] (Flow-CS)
  
  Details:
  Antibodies cited: 1. DR5 (IMG-120A): Flow (cell surface) 2. DR4 (IMG-141): Flow (cell surface, HCT116 cells), Supplementary Fig. 5. 3. DcR2 (IMG-121-1): WB (SW620 and HCT15 cells), Supplementary Fig. 4.
• Zhang X, Chen W, De Paiva C et al. Interferon exacerbates dry eye-induced apoptosis in conjunctiva through dual apoptotic pathways Invest Ophthalmol Vis Sci 2011-08-09 [PMID: 21474767] (ICC/IF, IF/IHC, Mouse)
• Jane EP, Premkumar DR, Pollack IF. Bortezomib sensitizes malignant human glioma cells to TRAIL, mediated by inhibition of the NF-{kappa}B signaling pathway. Mol Cancer Ther. 2011-01-01 [PMID: 21220502] (WB)
  
  Details:
  DR4 (IMG-141A) and DR5/TRAILR2 (IMG-120A). WB: Various human glioma cell lines treated with TRAIL: LN18, U87, LNZ308, and U373, Fig 2B & 2C.
• Wang S, Ren W, Liu J et al. TRAIL and doxorubicin combination induces proapoptotic and antiangiogenic effects in soft tissue sarcoma in vivo. Clin Cancer Res. 2010-05-01 [PMID: 20406839] (IHC-P, Human)
  
  Details:
  DR5/TRAILR2 (IMG-120A): IHC (paraffin), Human STS cell lines HT1080 and SkLMS1, Fig 1 and 2.
• Xu J, Zhou JY, Wei WZ et al. Sp1-mediated TRAIL induction in chemosensitization. Cancer Res. 2008-08-15 [PMID: 18701496] (WB, Human)
  
  Details:
  WB (human breast cancer MCF7 cells), Fig. 1A, C, D.
• Kusaba M, Nakao K, Goto T et al. Abrogation of constitutive STAT3 activity sensitizes human hepatoma cells to TRAIL-mediated apoptosis. J Hepatol. 2007-10-01 [PMID: 17602782]
• Xu J, Zhou JY, Wu GS. Tumor necrosis factor-related apoptosis-inducing ligand is required for tumor necrosis factor alpha-mediated sensitization of human breast cancer cells to chemotherapy. Cancer Res. 2006-10-15 [PMID: 17047073]
• Song JH, Bellail A, Tse MC et al. Human astrocytes are resistant to Fas ligand and tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis. J Neurosci. 2006-03-22 [PMID: 16554480]
• Younes M, Georgakis GV, Rahmani M et al. Functional expression of TRAIL receptors TRAIL-R1 and TRAIL-R2 in esophageal adenocarcinoma. Eur J Cancer. 2006-03-01 [PMID: 16426839] (IHC-P, WB, Flow-CS)
  
  Details:
  IHC-paraffin: formalin-fixed, paraffin-embedded tissue sections and cell line), (WB: cell lines), (Flow-cell surface: cell lines).
• Abdelrahim M, Newman K, Vanderlaag K et al. 3,3'-diindolylmethane (DIM) and its derivatives induce apoptosis in pancreatic cancer cells through endoplasmic reticulum stress-dependent upregulation of DR5. Carcinogenesis. 2006-04-01 [PMID: 16332727] (WB)
  
  Details:
  ATF6 (IMG-273) 2. DR5 (IMG-120) [WB, Fig.4 and Fig. 6 (Panc-28 cells)].
• Georgakis GV, Li Y, Humphreys R et al. Activity of selective fully human agonistic antibodies to the TRAIL death receptors TRAIL-R1 and TRAIL-R2 in primary and cultured lymphoma cells: induction of apoptosis and enhancement of doxorubicin- and bortezomib-induced cell death. Br J Haematol. 2005-08-01 [PMID: 16098063] (ICC/IF)
  
  Details:
  ICC: cytospins.
• Zou W, Liu X, Yue P et al. c-Jun NH2-terminal kinase-mediated up-regulation of death receptor 5 contributes to induction of apoptosis by the novel synthetic triterpenoid methyl-2-cyano-3,12-dioxooleana-1, 9-dien-28-oate in human lung cancer cells. Cancer Res. 2004-10-15 [PMID: 15492284]
• Fan QL, Zou WY, Song LH et al. Synergistic antitumor activity of TRAIL combined with chemotherapeutic agents in A549 cell lines in vitro and in vivo. Cancer Chemother Pharmacol. 2005-02-01 [PMID: 15290100]
• Jin Z, McDonald ER 3rd, Dicker DT et al. Deficient tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor transport to the cell surface in human colon cancer cells selected for resistance to TRAIL-induced apoptosis. J Biol Chem [PMID: 15155747]
  
  Details:
  Used the Azide Free form of this antibody.
• Dandrea T, Hellmold H, Jonsson C et al. The transcriptosomal response of human A549 lung cells to a hydrogen peroxide-generating system: relationship to DNA damage, cell cycle arrest, and caspase activation. Free Radic Biol Med [PMID: 15019973]
  
  Details:
  Used the Azide Free form of this antibody.
• Kim K, Nakagawa H, Fei P et al. Targeting Bcl-xL in esophageal squamous cancer to sensitize to chemotherapy plus TRAIL-induced apoptosis while normal epithelial cells are protected by blockade of caspase 9. Cell Death Differ [PMID: 14752513]
  
  Details:
  Used the Azide Free form of this antibody.
• Koyama S. Flow cytometric measurement of tumor necrosis factor-related apoptosis-inducing ligand and its receptors in gastric epithelium and infiltrating mucosal lymphocytes in Helicobacter pylori-associated gastritis. J Gastroenterol Hepatol. 2003-07-01 [PMID: 12795746] (Flow-CS)
• Kandasamy K, Srinivasula SM, Alnemri ES et al. Involvement of proapoptotic molecules Bax and Bak in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced mitochondrial disruption and apoptosis: differential regulation of cytochrome c and Smac/DIABLO release. Cancer Res [PMID: 12670926]
  
  Details:
  Used the Azide Free form of this antibody.
• Eid MA, Lewis RW, Kumar MV. Mifepristone pretreatment overcomes resistance of prostate cancer cells to tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL). Mol Cancer Ther. [PMID: 12492116]
  
  Details:
  Citation using the Azide Free form of this antibody.
• Zhang HG, Xie J, Xu L et al. Hepatic DR5 induces apoptosis and limits adenovirus gene therapy product expression in the liver. J Virol. 2002-06-01 [PMID: 11991997]
• Koyama S, Koike N, Adachi S. Expression of TNF-related apoptosis-inducing ligand (TRAIL) and its receptors in gastric carcinoma and tumor-infiltrating lymphocytes: a possible mechanism of immune evasion of the tumor. J Cancer Res Clin Oncol [PMID: 11862476]
  
  Details:
  Used the Azide Free form of this antibody.
• Burns TF, Bernhard EJ, El-Deiry WS. Tissue specific expression of p53 target genes suggests a key role for KILLER/DR5 in p53-dependent apoptosis in vivo. Oncogene. 2001-08-02 [PMID: 11498783]
• Cuello M, Ettenberg SA, Clark AS et al. Down-regulation of the erbB-2 receptor by trastuzumab (herceptin) enhances tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in breast and ovarian cancer cell lines that overexpress erbB-2. Cancer Res [PMID: 11406568]
  
  Details:
  Used the Azide Free form of this antibody.
• Basile JR, Zacny V, Munger K. The cytokines tumor necrosis factor-alpha (TNF-alpha ) and TNF-related apoptosis-inducing ligand differentially modulate proliferation and apoptotic pathways in human keratinocytes expressing the human papillomavirus-16 E7 oncoprotein. J Biol Chem. 2001-06-22 [PMID: 11306566] (WB, Human)
  
  Details:
  WB DR5/Apo2/TRAIL-R2/TRAILR2/TRICK2/KILLER (IMG-120A) and DcR2/TRAIL-R4/TRUNDD (IMG-121). Human keratinocytes (HFK) cells expressing HPV-16 EF (human papillomavirus), Fig 6.
• Kim K, Fisher MJ, Xu SQ, el-Deiry WS. Molecular determinants of response to TRAIL in killing of normal and cancer cells. Clin Cancer Res. 2000-02-01 [PMID: 10690508]

Review for TRAILR2/TNFRSF10B Antibody (NB100-56618) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 07/17/2018

Summary

• Application: Western Blot
• Sample Tested: Breast cancer cells
• Species: Human
• Lot: 0716499c-04

Comments

• Comments: Total cell lysates from T47D and MDA-MB-231 cells were subjected to western blot. PVDF membrane were probed with 2.5 mm/ml Human TRAIL R2 (NB100-56618). A specific band was detected for TRAIL R2 at approximately 48~50 kDa. This experiment was conducted under reducing conditions

Bioinformatics

• Gene Symbol: TNFRSF10B
• Entrez:
  • 8795(Human)