DR5 Antibody

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ProSci
Product Code: PSI-2019
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2019-0.02mg0.02mg£150.00
Quantity:
PSI-2019-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunofluorescence (IF)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 2 Western Blot Validation in Human HepG2 Cells</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 μg/mL Lane 2: 2 μg/mL Lane 3: 4 μg/mL
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<strong>Figure 3 Western Blot Validation in Mouse and Rat Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP
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<strong>Figure 4 Western Blot Validation in Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 5 Western Blot Validation in Mouse Heart </strong><br> Loading: 15 μg of lysatesper lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 6 Western Blot Validation in Rat Skeletal Muscle </strong><br> Loading: 15 μg of lysate per lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 7 Immunofluorescence Validation of DR5 in Human HepG2 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human HepG2 cells labeling DR5 with 2019 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
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<strong>Figure 8 Immunofluorescence Validation of DR5 in Human Testis </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling DR5 with 2019 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
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<strong>Figure 9 Immunofluorescence Validation of DR5 in Mouse Pancreas </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas tissue labeling DR5 with 2019 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
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<strong>Figure 10 Immunofluorescence Validation of DR5 in Rat Brain </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed rat brain tissue labeling DR5 with 2019 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
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<strong>Figure 11 Immunohistochemistry Validation of DR5 in Mouse kidney tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-DR5 antibody (2019) at 5μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 12 KO Validation of DR5 in HCT116 Cells (Han et al., 2015) </strong><br> Anti-cancer drug, Carfilzomib (CFZ), induced up-regulation of DR5 and the expression of DR5 was not detected in DR5-KO HCT 116 cell line with anti-DR5 antibodies (2019).
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<strong>Figure 13 KD Validation of DR5 in MB231 Cells (Rahman et al., 2009) </strong><br> Western blot analysis with anti-DR5 antibodies was performed for DR5 in MB231 cells transfected with control siRNA or DR5 siRNA. DR5 expression was disrupted after DR5 siRNA knockdown.
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<strong>Figure 14 Immunohistochemistry Validation of BIM in Human Colon Tumors (Devetzi et al., 2016) </strong><br> Protein analysis for DR5 by immunohistochemistry with anti-DR5 antibodies in human colon tumors. Strong immunoreactivity is shown for DR5 in T167 patient with colorectal cancer.
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<strong>Figure 15 Regulated Expression Validation of DR5 in Thyroid Epithelial Cells (Bretz et al., 2002) </strong><br> Immunostaining with anti-DR5 antibodies shows high levels of DR5 expression in untreated cells and cells treated with each of the three cytokines alone or TNFalpha combined with IL-1b. In contrast, treatment with both IFNg and TNFalpha or all three cytokines greatly reduces DR5 staining. The reduction in staining appears most significant in cytoplasmic regions while some staining is maintained in or around the nucleus.
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<strong>Figure 1 Western Blot Validation in Human Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (0.5 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation in Human HepG2 Cells</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 μg/mL Lane 2: 2 μg/mL Lane 3: 4 μg/mL
<strong>Figure 3 Western Blot Validation in Mouse and Rat Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP
<strong>Figure 4 Western Blot Validation in Mouse Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Western Blot Validation in Mouse Heart </strong><br> Loading: 15 μg of lysatesper lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 6 Western Blot Validation in Rat Skeletal Muscle </strong><br> Loading: 15 μg of lysate per lane. Antibodies: DR5 2019, (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 7 Immunofluorescence Validation of DR5 in Human HepG2 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human HepG2 cells labeling DR5 with 2019 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
<strong>Figure 8 Immunofluorescence Validation of DR5 in Human Testis </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling DR5 with 2019 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
<strong>Figure 9 Immunofluorescence Validation of DR5 in Mouse Pancreas </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse pancreas tissue labeling DR5 with 2019 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
<strong>Figure 10 Immunofluorescence Validation of DR5 in Rat Brain </strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed rat brain tissue labeling DR5 with 2019 at 5 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI (blue).
<strong>Figure 11 Immunohistochemistry Validation of DR5 in Mouse kidney tissue</strong><br> Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti-DR5 antibody (2019) at 5μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 12 KO Validation of DR5 in HCT116 Cells (Han et al., 2015) </strong><br> Anti-cancer drug, Carfilzomib (CFZ), induced up-regulation of DR5 and the expression of DR5 was not detected in DR5-KO HCT 116 cell line with anti-DR5 antibodies (2019).
<strong>Figure 13 KD Validation of DR5 in MB231 Cells (Rahman et al., 2009) </strong><br> Western blot analysis with anti-DR5 antibodies was performed for DR5 in MB231 cells transfected with control siRNA or DR5 siRNA. DR5 expression was disrupted after DR5 siRNA knockdown.
<strong>Figure 14 Immunohistochemistry Validation of BIM in Human Colon Tumors (Devetzi et al., 2016) </strong><br> Protein analysis for DR5 by immunohistochemistry with anti-DR5 antibodies in human colon tumors. Strong immunoreactivity is shown for DR5 in T167 patient with colorectal cancer.
<strong>Figure 15 Regulated Expression Validation of DR5 in Thyroid Epithelial Cells (Bretz et al., 2002) </strong><br> Immunostaining with anti-DR5 antibodies shows high levels of DR5 expression in untreated cells and cells treated with each of the three cytokines alone or TNFalpha combined with IL-1b. In contrast, treatment with both IFNg and TNFalpha or all three cytokines greatly reduces DR5 staining. The reduction in staining appears most significant in cytoplasmic regions while some staining is maintained in or around the nucleus.

Documents

Further Information

Additional Names:
DR5 Antibody: DR5, CD262, KILLER, TRICK2, TRICKB, ZTNFR9, TRAILR2, TRICK2A, TRICK2B, TRAIL-R2, KILLER/DR5, DR5, UNQ160/PRO186, Tumor necrosis factor receptor superfamily member 10B, Death receptor 5, TRAIL receptor 2
Application Note:
WB: 0.5-2 μg/mL; IHC-P: 5 μg/mL; IF: 5-10 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in mouse samples; Immunocytochemistry in human samples and Immunofluorescence in human, mouse and rat samples. All other applications and species not yet tested.
Background:
DR5 Antibody: 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. 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-κB.
Background References:
  • Pan et al. Science 1997;277:815-8.
  • Sheridan et al. Science 1997;277:818-21.
  • Walczak? et al. EMBO J 1997;16:5386-97.
  • MacFarlane et al. J Biol Chem 1997;272:25417-20
Buffer:
DR5 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/ml
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Immunogen:
DR5 antibody was raised against a peptide corresponding to 20 amino acids near the carboxy terminus of human DR5 precursor.

The immunogen is located within the last 50 amino acids of DR5.
ISOFORMS:
Human DR5 has 3 isoforms, including isoform L (440aa, 48kD), isoform S (411aa, 45kD) and isoform 3 (118aa, 13kD). Mouse DR5 has 1 isoform (381aa, 42kD), and Rat DR5 also has 1 isoform (336aa, 37kD). 2019 can detect two human isoforms and can also detect mouse and rat isoforms.
NCBI Gene ID #:
8795
NCBI Official Name:
tumor necrosis factor receptor superfamily, member 10b
NCBI Official Symbol:
TNFRSF10B
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 48kD and 45kD

Observed: 48kD and 45kD
Protein Accession #:
AF012535
Protein GI Number:
2338419
Purification:
DR5 Antibody is affinity chromatography purified via peptide column.
Research Area:
Apoptosis
SPECIFICITY:
Antibody has no cross reaction to DR4.
Swissprot #:
O14763
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

KO Validation (Figure 12) shows DR5 expression detected by anti-DR5 antibodies (2019) was disrupted in DR5 KO HCT116 cells. 

KD validation (Figure 13): Anti-DR5 antibody (2019) specificity was further verified by DR5 specific knockdown. DR5 signal in MB231 cells transfected with DR5 siRNAs was disrupted in comparison with that in cells transfected with control siRNAs.

Regulated expression validation (Figure 12, 15): DR5 expression detected by anit-DR5 antibodies (2019) was up-regulated by carfilzomib treatment and was down-regulated by treatment with two or more cytokines.

References

  1. Han et al. The novel proteasome inhibitor carfilzomib activates and enhances extrinsic apoptosiS involving stabilization of death receptor 5. Oncotarget. 2015;6(19):17532-42. PMID: 26009898
  2. Rahman et al. TRAIL induces apoptosis in triple-negative breast cancer cells with a mesenchymal phenotype. Breast Cancer Res Treat. 2009;113(2):217-30. doi:10.1007/s10549-008-9924-5. Epub 2008.PMID: 18266105
  3. Devetzi et al. Death receptor 5 (DR5) and a 5-gene apoptotic biomarker panel with significant differential diagnostic potential in colorectal cancer. Sci Rep. 2016 ;6:36532. PMID: 27827395
  4. Bretz et al.Inflammatory cytokine regulation of TRAIL-mediated apoptosis in thyroid epithelial cells. Cell Death Differ. 2002;9(3):274-86.PMID: 11859410
  5. Candolfi et al. Release of HMGB1 in response to proapoptotic glioma killing strategies: efficacy and neurotoxicity. Clin Cancer Res. 2009;15(13):4401-14. doi: 10.1158/1078-0432.CCR-09-0155. PMID: 19570774
  6. Malhi et al. Free fatty acids sensitise hepatocytes to TRAIL mediated cytotoxicity. Gut. 2007;56(8):1124-31. PMID: 17470478
  7. Razumilava et al. miR-25 targets TNF-related apoptosis inducing ligand (TRAIL) death receptor-4 and promotes apoptosis resistance in cholangiocarcinoma. Hepatology. 2012;55(2):465-75.PMID: 21953056
  8. Koyama et al. Histone deacetylase inhibitors and 15-deoxy-Delta12,14-prostaglandin J2 synergistically induce apoptosis. Clin Cancer Res. 2010;16(8):2320-32. PMID: 20371690
  9. Kang et al. Drozitumab, a human antibody to death receptor 5, has potent antitumor activity against rhabdomyosarcoma with the expression of caspase-8 predictive of response. Clin Cancer Res. 2011;17(10):3181-92. doi: 10.1158/1078-0432.CCR-10-2874. Epub 2011.PMID: 21385927
  10. Gupta et al. Nimbolide, a limonoid triterpene, inhibits growth of human colorectal cancer xenografts by suppressing the proinflammatory microenvironment. Clin Cancer Res. 2013;19(16):4465-76. PMID: 23766363
  11. Yuan et al. PARP-1 regulates resistance of pancreatic cancer to TRAIL therapy. Clin Cancer Res. 2013;19(17):4750-9.PMID: 23833311
  12. Zhao et al. mTOR complex 2 is involved in regulation of Cbl-dependent c-FLIP degradation and sensitivity of TRAIL-induced apoptosis. Cancer Res. 2013;73(6):1946-57. PMID: 23319802
  13. Dhandapani et al. Retinoic acid enhances TRAIL-induced apoptosis in cancer cells by upregulating TRAIL receptor 1 expression. Cancer Res. 2011;71(15):5245-54. PMID: 21685476
  14. Sun et al. Identification of an antiapoptotic protein complex at death receptors. Cell Death Differ. 2008;15(12):1887-900. PMID: 18846110
  15. Kurita et al. GLI3-dependent repression of DR4 mediates hedgehog antagonism of TRAIL-induced apoptosis. Oncogene. 2010;29(34):4848-58. PMID: 20562908

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