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Immunofluorescence analysis of TRAIL-R2/DR5 was done on 70% confluent log phase HeLa cell. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TRAIL-R2/Rabbit Polyclonal Antibody (Product # PA1-957) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
Western blot analysis was performed on whole cell extracts (20 µg lysate) of A549 (Lane 1) and HEL 92.1.7 (Lane 2). The blots were probed with Anti-TRAIL-R2 Rabbit Polyclonal Antibody (Product # PA1-957, 0.5-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 58 kDa band corresponding to TRAIL-R2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Western blot analysis of TRAIL Receptor 2 was performed by loading 25 ug of Hela (lane 1) and PC12 (lane 2) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a TRAIL Receptor 2 polyclonal antibody (Product # PA1-957) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~58 kDa in Hela cells.
Western Blot (WB)
Host / Isotype
Synthetic Peptide: C V(255) P E Q E M E V Q E P A E P T G(270)
Antigen affinity chromatography
PBS with 1mg/ml BSA
0.05% sodium azide
-20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
PA1-957 detects death receptor 5 (DR5) from human and rat tissues and cells.
PA1-957 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~58 kDa protein representing DR5 from HEK293 cell lysate. PA1-957 immunizing peptide corresponds to amino acid residues 255-270 of rat DR5. PA1-957 immunizing peptide (Cat. # PEP-048) is available for use in neutralization and control experiments.
DR5 is a recently identified death domain containing receptor for TRAIL, which mediates TRAIL induced apoptosis. DR5 is also known as Apo2, TRAIL-R2, TRICK2 and KILLER.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Protein Aliases: apoptosis inducing protein TRICK2A/2B; apoptosis inducing receptor TRAIL-R2; CD262; cytotoxic TRAIL receptor-2; death domain containing receptor for TRAIL/Apo-2L; Death receptor 5; DR5; Fas-like protein; KILLER; KILLER/DR5; p53-regulated DNA damage-inducible cell death receptor(killer); TNF-related apoptosis-inducing ligand receptor 2; TRAIL receptor 2; TRAIL-R2; TRAILR2; TRICK2; TRICK2A; TRICK2B; TRICKB; Tumor necrosis factor receptor superfamily member 10B; tumor necrosis factor receptor superfamily, member 10b; tumor necrosis factor receptor-like protein ZTNFR9; ZTNFR9
Gene Aliases: CD262; DR5; KILLER; KILLER/DR5; TNFRSF10B; TRAIL-R2; TRAILR2; TRICK2; TRICK2A; TRICK2B; TRICKB; UNQ160/PRO186; ZTNFR9
UniProt ID: (Human) O14763
Entrez Gene ID: (Human) 8795, (Rat) 364420
tumor necrosis factor receptor
Suggested Secondary Antibodies
Material safety data sheets (MSDS)