NF-kappaB p65 (phospho-T435) antibody

• Catalog Number: orb304548
• Category: Antibodies
• Description: Rabbit polyclonal antibody to NF-kappaB p65 (phospho-T435)
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IF, IH, IP, WB
• Reactivity: Human, Mouse, Primate, Rat
• Immunogen: KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding T435 of human NF-kappaB p65 protein. The exact sequence is proprietary.
• Dilution range: WB: 1:500-1000, IHC-P: 1:100-200, IF/ICC: 1:100-500, IP: 1:10-100
• Form/Appearance: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
• Conjugation: Unconjugated
• Target: RELA
• Entrez: 19697, 5970
• UniProt ID: Q04207, Q04206
• Source: Rabbit
• Storage: Shipped at 4°C. Upon delivery aliquot and store at -20°C for one year. Avoid freeze/thaw cycles.
• Buffer/Preservatives: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
• Alternative names: anti NFKB3 antibody, anti Transcription factor p65
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western blot analysis of NF-kappaB p65 (Phospho-T435) expression in Jurkat TNFa-treated (A), HepG2 TNFa-treated (B) whole cell lysates. (Predicted band size: 60 kD; Observed band size: 75 kD)

Immunohistochemical analysis of NF-kappaB p65 (Phospho-T435) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (Phospho-H 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of NF-kappaB p65 (Phospho-T435) staining in K562 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark.

Direct ELISA antibody dose-response curve using Anti-NF-kappaB p65 (Phospho-T435) Antibody. Antigen (Phosphopeptide and non-phosphopeptide) concentration is 5 ug/ml. Goat Anti-Rabbit IgG (H&L) - HRP was used as the secondary antibody, and signal was developed by TMB substrate.