Overview

  • Product name

    Anti-DR5 antibody [EPR19310]
    See all DR5 primary antibodies
  • Description

    Rabbit monoclonal [EPR19310] to DR5
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human DR5 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: O14763

  • Positive control

    • WB: Untreated and treated with 0.5µM/ml doxorubicin for 24 hours HCT 116 whole cell lysates; HeLa, HepG2 and HT1080 whole cell lysates; Human melanoma lysate. ICC/IF: HT1080 and HCT 116 cells. Flow Cyt: HCT 116 cells. IP: HT-1080 treated with 5µM MG132 for 4 hour whole cell lysate; HCT 116 whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199357 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 48, 40 kDa (predicted molecular weight: 48 kDa).
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/70.

Target

  • Function

    Receptor for the cytotoxic ligand TNFSF10/TRAIL. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. Promotes the activation of NF-kappa-B. Essential for ER stress-induced apoptosis.
  • Tissue specificity

    Widely expressed in adult and fetal tissues; very highly expressed in tumor cell lines such as HeLaS3, K-562, HL-60, SW480, A-549 and G-361; highly expressed in heart, peripheral blood lymphocytes, liver, pancreas, spleen, thymus, prostate, ovary, uterus, placenta, testis, esophagus, stomach and throughout the intestinal tract; not detectable in brain.
  • Involvement in disease

    Squamous cell carcinoma of the head and neck
  • Sequence similarities

    Contains 1 death domain.
    Contains 3 TNFR-Cys repeats.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Fas like protein antibody
    • Apoptosis inducing protein TRICK2A/2B antibody
    • Apoptosis inducing receptor TRAIL R2 antibody
    • CD262 antibody
    • CD262 antigen antibody
    • Cytotoxic TRAIL receptor 2 antibody
    • Death domain containing receptor for TRAIL/Apo 2L antibody
    • Death receptor 5 antibody
    • DR5 antibody
    • KILLER antibody
    • KILLER/DR5 antibody
    • OTTHUMP00000123492 antibody
    • OTTHUMP00000123493 antibody
    • p53 regulated DNA damage inducible cell death receptor(killer) antibody
    • TNF related apoptosis inducing ligand receptor 2 antibody
    • TNF-related apoptosis-inducing ligand receptor 2 antibody
    • TNFRSF10B antibody
    • TR10B_HUMAN antibody
    • TRAIL R2 antibody
    • TRAIL receptor 2 antibody
    • TRAIL-R2 antibody
    • TRAILR2 antibody
    • TRICK2 antibody
    • TRICK2A antibody
    • TRICK2B antibody
    • TRICKB antibody
    • Tumor necrosis factor receptor like protein ZTNFR9 antibody
    • Tumor necrosis factor receptor superfamily member 10B antibody
    • Tumor necrosis factor receptor superfamily, member 10b antibody
    • ZTNFR9 antibody
    see all

Images

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling DR5 with ab199357 at 1/70 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • All lanes : Anti-DR5 antibody [EPR19310] (ab199357) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : TNFRSF10B knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 48 kDa
    Observed band size: 47 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 2: Merged signal (red and green). Green - ab199357 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab199357 was shown to recognize DR5 in wild-type HAP1 cells as signal was lost at the expected MW in TNFRSF10B knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and TNFRSF10B knockout samples were subjected to SDS-PAGE. Ab199357 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT1080 (Human fibrosarcoma cell line) cells labeling DR5 with ab199357 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HT1080  cells. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab199357 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • All lanes : Anti-DR5 antibody [EPR19310] (ab199357) at 1/1000 dilution

    Lane 1 : Untreated HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 2 : HCT 116 (Human colorectal carcinoma cell line) treated with 0.5µM/ml doxorubicin for 24 hours whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48 kDa
    Observed band size: 40,48 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Doxorubicin treatment elevated the expression of DR5 (PMID: 12496481; PMID: 11468181; PMID: 11090076). The expression profile is consistent with what has been described in the literature (PMID:20515924; PMID:16297203).

  • All lanes : Anti-DR5 antibody [EPR19310] (ab199357) at 1/1000 dilution

    Lane 1 : Human melanoma lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 4 : HT1080 (Human fibrosarcoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 48 kDa
    Observed band size: 40,48 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-2: 3 minutes; Lane 3: 10 seconds; Lane 4: 8 seconds.

    The expression profile is consistent with what has been described in the literature (PMID:20515924; PMID:16297203).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling DR5 with ab199357 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on HCT 116 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab199357 at 1/100 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • DR5 was immunoprecipitated from 0.35mg of HT1080 (Human fibrosarcoma cell line) treated with 5μM MG132 for 4 hour whole cell lysate with ab199357 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199357 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HT1080 treated with 5μM MG132 for 4 hour whole cell lysate, 10µg (Input).

    Lane 2: ab199357 IP in HT1080 treated with 5μM MG132 for 4 hour whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab199357 in HT1080 treated with 5μM MG132 for 4 hour whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • DR5 was immunoprecipitated from 0.35mg of HCT 116 (Human colorectal carcinoma cell line) whole cell lysate with ab199357 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199357 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HCT 116 whole cell lysate, 10µg (Input).

    Lane 2: ab199357 IP in HCT 116 whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730)  instead of ab199357 in HCT 116 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

This product has been referenced in:

  • Qian H  et al. Combined application of Embelin and tumor necrosis factor-related apoptosis-inducing ligand inhibits proliferation and invasion in osteosarcoma cells via caspase-induced apoptosis. Oncol Lett 15:6931-6940 (2018). Read more (PubMed: 29731867) »
  • Yao TH  et al. Propranolol induces hemangioma endothelial cell apoptosis via a p53-BAX mediated pathway. Mol Med Rep 18:684-694 (2018). Read more (PubMed: 29767244) »
See all 5 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (B16 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycine Gel, PVDF membrane)
Loading amount
40 µg
Specification
B16 cells
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 03 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (panc1 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycine Gel, PVDF membrane)
Loading amount
40 µg
Specification
panc1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 03 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (B16 tumors taken out of C57BL/6 mice)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
B16 tumors taken out of C57BL/6 mice
Blocking step
Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100%
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Flow Cytometry
Sample
Human Cell (MDA-MB-231 cells)
Permeabilization
No
Gating Strategy
no gating
Specification
MDA-MB-231 cells
Preparation
Cell harvesting/tissue preparation method: Cells scrapped in PBS
Sample buffer: PBS with 0.5% BSA
Fixation
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 12 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (MDA-MB-231 tumors taken out Nude Balb/C mice)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
MDA-MB-231 tumors taken out Nude Balb/C mice
Blocking step
Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 18 2017

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