DR4 Antibody

• Catalog Number: orb1239340
• Category: Antibodies
• Description: DR4 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Anti-DR4 antibody (orb1239340) was raised against a peptide corresponding to 19 amino acids near the carboxy terminus of human DR4. The immunogen is located within the last 50 amino acids of DR4.
• Antibody Type: Primary Antibody
• Concentration: 1 mg/mL
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 50kDObserved: 55kD (Post-modification: 1 N-linked glycosylation)
• Target: TNFRSF10A
• UniProt ID: O00220
• NCBI: AAC51226
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: DR4 Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: DR4 Antibody: DR4, APO2, CD261, TRAILR1, TRAILR-1,
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

KO Validation in HeLa Cells. Loading: 10 µg of HeLa WT cell lysates or DR4 KO cell lysates. Antibodies: DR4 orb1239340 (1 µg/mL) and beta-actin orb1240312 (1 µg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: DR4 orb1239340 (1 µg/mL), DR4 orb1239343 (4 µg/mL), and beta-actin (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Cell Lines. Loading: 15 µg of cell lysates per lane. Antibodies: DR4 orb1239340 (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of DR4. Immunofluorescent analysis of 4% paraformaldehyde-fixed human spleen tissue labeling DR4 with orb1239340 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue). Image showing membrane staining on human spleen cells.

Immunocytochemistry Validation of DR4 in HeLa Cells. Immunocytochemical analysis of HeLa cells using anti-DR4 antibody (orb1239340) at 10 µg/ml. Cells 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.

Immunohistochemistry Validation of DR4. Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-DR4 antibody (orb1239340) at 10 µg/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.

KD Validation in SW480 cells (Goda et al., 2008). The expression of DR4 was knocked down via DR4 siRNA, 24 h latercells were treated with dipyridamole for 24 h. DR4 protein expression detected by anti-DR4 antibodies (orb1239340) was disrupted. Dipyridamole up-regulated the expression of DR4.

Immunofluorescence Validation of DR4 in rat brain (Cantarella et al., 2014). DR4 protein expression detected by anti-DR4 antibodies (orb1239340) was increased after transient brain ischemia (tMCAO) and decreased after pre-conditioning stimulus. Confocal microscopic images displaying NeuN (a, d, g) (green), DR4 (b, e, h) (red), and Merge (c, f, i) (yellow) in the brain peri-ischemic region of rats after 5 h.

Immunocytochemistry Validation of DR4 in human melanoma cells (Ekmekcioglu et al., 2008). MeWo melanoma cells were exposed to affinity-purified MDA7/IL-24. After 48 h of treatment, cells were collected and cytospins prepared for cytochemical assessment of their TRAIL receptor (R1 and R2) expression (anti-DR4 (orb1239340) or anti-DR5, AEC, hematoxylin). Both DR4 and DR5 expression were upregulated in MeWo cells after treatment.

KD Validation in Huh7 cells (Malhi et al., 2007). Western blot analysis with anti-DR4 antibodies (orb1239340) was performed for DR5 and DR4 expression using whole cell lysates from Huh 7 cells transfected with respective siRNAs. In cells treated with siDR4, a decrease in DR4 level was observed, DR5 levels were unchanged. Scrambled siRNA was used as control.

KD Validation in HeLa cells (Horinaka et al., 2005). HeLa cells were transfected with DR4siRNA or LacZ control siRNA. At 24 h after transfection, the cells were treated with or without 20 µM luteolin for 24 h. Western blot analysis was carried out with anti-DR4 antibodies (orb1239340). DR4 expression was markedly reduced after DR4 knockdown.