NF-κB p6/RELA/NF Rabbit Polyclonal Antibody

SKU: orb1939972

Description

Anti-NF-κB p65/RELA Antibody. Tested in ELISA, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Research Area

Signal Transduction

Images & Validation

• Tested Applications: ELISA, FC, WB
• Dilution Range: Western blot, 0.1-0.25 μg/ml, Human, Mouse, Rat Flow Cytometry (Fixed), 1-3 μg/1x10^6 cells, Human ELISA, 0.1-0.5 μg/ml, -
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: E.coli-derived human NF-κB p65/RELA recombinant protein (Position: Q132-S551). Human RELA shares 85.8% amino acid (aa) sequence identity with mouse RELA.
• Target: Transcription factor p65
• Molecular Weight: 65 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

NF Kappa B; NF κB; NFKB 3; NFKB p65; NFKB3; NF-κB; NF-κB3; p65; RELA; Transcription factor p65

Flow Cytometry analysis of Daudi cells using anti-NF-ΚB P6/RELA antibody. Overlay histogram showing Daudi cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NF-ΚB P6/RELA Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Western blot analysis of NF-ΚB P6/RELA using anti-NF-ΚB P6/RELA antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human Caco-2 whole cell lysates, Lane 4: human Raji whole cell lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: rat C6 whole cell tissue lysates, Lane 7: mouse NIH/3T3 whole cell lysates, Lane 8: mouse Ana-1 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NF-ΚB P6/RELA antigen affinity purified polyclonal antibody at 0.25 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NF-ΚB P6/RELA at approximately 65 kDa. The expected band size for NF-ΚB P6/RELA is at 60 kDa.

Quick Database Links

Gene Symbol

Transcription factor p65

UniProt

Q04206