Human TRAIL R2/TNFRSF10B PE‑conjugated Antibody

Detection of TRAIL R2/TNFRSF10B by Western Blot

Knockdown of K8 enhances TRAIL induced apoptosis(A-C) Cells were transfected with a control siRNA or siRNA specific to KRT8 transcript for 72 hours, followed by TRAIL stimulation (100 ng/ml [T47D and BT474] or 150 ng/ml [MCF7]) for 24 hours. The resultant cells were analyzed by immunoblotting using antibodies specific to K8/K18, DR4, DR5, caspase-3, caspase-8, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Caspase activation was identified by decrease in pro-enzyme form (ProC-8 and ProC-3). (D-F) Cells were treated as above and analyzed by flow cytometry after staining with Annexin-V-FITC and propidium iodide (PI). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29796187), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TRAIL R2/TNFRSF10B by Flow Cytometry

K8/K18 physically interacts with DR5(A) Cell lysates from MCF7 and T47D cell lines were subjected to immunoprecipitation (IP) with an anti-K8/K18 antibody immobilized to protein A-agarose (lane 2 and 4) or with protein A-agarose alone (lane 1 and 3). IP samples were immunoblotted for DR5. (B) Cells were fixed, permeabilized, stained with anti-DR5 antibody (red) and anti-keratin 8 antibody (green), and analyzed by confocal microscopy. All images were acquired using a 40x objective lens. (C) MCF7 cells were transfected with control siRNA or siRNA against KRT8 for 72 hr. The resultant cells were fixed, permeabilized, and stained with anti-DR5 antibody (red), DAPI (nuclei, blue), and fluorescent phalloidin 488 for actin visualization (green). All images were acquired using a 40x objective lens. (D-F) Flow cytometry analysis of cells transfected with control siRNA or siRNA against KRT8. The resultant cells were harvested using a non-enzymatic cell dissociation solution. The cell samples were incubated with PE conjugated anti-DR5 antibody (IgG2b) or its corresponding IgG2b isotype as a control. (D) Shown are representative histograms of unstained cells (black dashed line), cells stained with the isotype control (red fill), and cells stained with PE-anti-DR5 antibody (blue outline with no fill). The right shift represents the presence of DR5 on the cell surface membrane. Median PE-anti-DR5 antibody fluorescence intensity for cells transfected with control siRNA and siRNA targeting keratin 8 are shown after subtraction of isotype control median fluorescence intensity in two independent experiments for T47D cells (E) and MCF7 cells (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29796187), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TRAIL R2/TNFRSF10B by Immunocytochemistry/ Immunofluorescence

K8/K18 physically interacts with DR5(A) Cell lysates from MCF7 and T47D cell lines were subjected to immunoprecipitation (IP) with an anti-K8/K18 antibody immobilized to protein A-agarose (lane 2 and 4) or with protein A-agarose alone (lane 1 and 3). IP samples were immunoblotted for DR5. (B) Cells were fixed, permeabilized, stained with anti-DR5 antibody (red) and anti-keratin 8 antibody (green), and analyzed by confocal microscopy. All images were acquired using a 40x objective lens. (C) MCF7 cells were transfected with control siRNA or siRNA against KRT8 for 72 hr. The resultant cells were fixed, permeabilized, and stained with anti-DR5 antibody (red), DAPI (nuclei, blue), and fluorescent phalloidin 488 for actin visualization (green). All images were acquired using a 40x objective lens. (D-F) Flow cytometry analysis of cells transfected with control siRNA or siRNA against KRT8. The resultant cells were harvested using a non-enzymatic cell dissociation solution. The cell samples were incubated with PE conjugated anti-DR5 antibody (IgG2b) or its corresponding IgG2b isotype as a control. (D) Shown are representative histograms of unstained cells (black dashed line), cells stained with the isotype control (red fill), and cells stained with PE-anti-DR5 antibody (blue outline with no fill). The right shift represents the presence of DR5 on the cell surface membrane. Median PE-anti-DR5 antibody fluorescence intensity for cells transfected with control siRNA and siRNA targeting keratin 8 are shown after subtraction of isotype control median fluorescence intensity in two independent experiments for T47D cells (E) and MCF7 cells (F). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29796187), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TRAIL R2/TNFRSF10B by Western Blot

Knockdown of K8 enhances TRAIL induced apoptosis(A-C) Cells were transfected with a control siRNA or siRNA specific to KRT8 transcript for 72 hours, followed by TRAIL stimulation (100 ng/ml [T47D and BT474] or 150 ng/ml [MCF7]) for 24 hours. The resultant cells were analyzed by immunoblotting using antibodies specific to K8/K18, DR4, DR5, caspase-3, caspase-8, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Caspase activation was identified by decrease in pro-enzyme form (ProC-8 and ProC-3). (D-F) Cells were treated as above and analyzed by flow cytometry after staining with Annexin-V-FITC and propidium iodide (PI). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29796187), licensed under a CC-BY license. Not internally tested by R&D Systems.