Anti-TNF alpha Antibody

Product Name

Anti-TNF alpha Antibody

View all TNF-alpha Antibodies

SKU/Catalog Number

PA1079

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-TNF alpha Antibody catalog # PA1079. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-TNF alpha Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1079)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

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.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1079 is reactive to TNF in Human, Mouse, Rat

Applications

PA1079 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

25 kDa

Calculated molecular weight

25.644kDa

Background of TNF-alpha

TNF alpha (Tumor Necrosis Factor alpha) gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, By Heat
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of TNF alpha using anti-TNF alpha antibody (PA1079).
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 (Catalog # PA1079) 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 (Catalog # EK1002) 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.

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Figure 2. IHC analysis of TNF alpha using anti-TNF alpha antibody (PA1079).
TNF alpha was detected in a paraffin-embedded section of human B lymphocytic tumor tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, 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 (PA1079) 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 (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of CACO-2 cells using anti-TNF alpha antibody (PA1079).
Overlay histogram showing CACO-2 cells stained with PA1079 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF alpha Antibody (PA1079, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For TNF (Source: Uniprot.org, NCBI)

Alternative Names

APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha TNF DIF-alpha, TNFA, TNFSF2, TNLG1F, TNF tumor necrosis factor tumor necrosis factor|APC1 protein|TNF, macrophage-derived|TNF, monocyte-derived|TNF-a|tumor necrosis factor ligand 1F|tumor necrosis factor ligand superfamily member 2|tumor necrosis factor-alpha

For more info on TNF, check out the TNF Infographic

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In this infographic, you will see the following information for TNF: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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