TNF alpha Rabbit Polyclonal Antibody

SKU: orb526980

Description

TNF alpha Rabbit Polyclonal Antibody

Research Area

Cancer Biology, Cardiovascular Research, Immunology & Inflammation, Metabolism Research, Signal Transduction

Images & Validation

• Tested Applications: FC, IHC, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x10^6 cells, Human
• Reactivity: Human, Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human TNF alpha, different from the related mouse sequence by five amino acids, and rat sequence by seven amino acids.
• Target: Non-structural protein 3
• Molecular Weight: 25 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 and 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

Cachectin; DIF; TNF; TNF a; TNF alpha; TNFA; TNF-alpha; TNFSF2; TNFα; TNF-α; Tumor necrosis factor

Flow Cytometry analysis of CACO-2 cells using anti-TNF alpha antibody. Overlay histogram showing CACO-2 cells (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF alpha 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.

IHC analysis of TNF alpha using anti-TNF alpha antibody. TNF alpha was detected in a paraffin-embedded section of human B lymphocytic tumor tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml rabbit anti-TNF alpha Antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Western blot analysis of TNF alpha using anti-TNF alpha 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 U937 whole cell lysates, Lane 2: rat spleen tissue lysates, Lane 3: rat C6 whole cell lysates, Lane 4: mouse spleen tissue 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-TNF alpha antigen affinity purified polyclonal antibody at 0.5 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 TNF alpha at approximately 25 kDa. The expected band size for TNF alpha is at 26 kDa.

Quick Database Links

Gene Symbol

Non-structural protein 3

UniProt

P01375