TNF alpha Rabbit Polyclonal Antibody

SKU: orb669133

Description

Anti-TNF alpha/Tnf Antibody. Tested in ELISA, Flow Cytometry, IF, ICC, and WB applications. This antibody reacts with Mouse, Rat, offering an essential tool for scientific applications such as cell lysate analysis and breast adenocarcinoma cell line research.

Research Area

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

Images & Validation

• Tested Applications: ELISA, WB
• Dilution Range: Western blot, 0.25-0.5μg/ml, Mouse, Rat ELISA, 0.1-0.5μg/ml, -
• Reactivity: Mouse, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse TNF alpha/Tnf recombinant protein (Position: D89-E185).
• Target: Mucin-3A/B
• Molecular Weight: 17 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: 500 µg/ml
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

Tumor necrosis factor; Cachectin; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; Tumor necrosis factor, membrane form; N-terminal fragment; NTF; Intracellular domain 1; ICD1; Intracellular domain 2; ICD2; C-domain 1; C-domain 2; Tumor necrosis factor, soluble form; Tnf; Tnfa, Tnfsf2

Flow Cytometry analysis of Raw264.7 cells using anti-TNF antibody. Overlay histogram showing Raw264.7 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of TNF Alpha/Tnf using anti-TNF Alpha/Tnf antibody. TNF Alpha/Tnf was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-TNF Alpha/Tnf Antibody overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of TNF Alpha/Tnf using anti-TNF Alpha/Tnf 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 50 ug of sample under reducing conditions. Lane 1: rat thymus tissue lysates, Lane 2: mouse thymus 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/Tnf 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/Tnf at approximately 25 KD. The expected band size for TNF Alpha/Tnf is at 25 KD.

Quick Database Links

Gene Symbol

Mucin-3A/B

UniProt

P06804