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Western Blot (WB) (Figure 1. 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.Lane 1: recombinant human TNF alpha protein 1ng.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 ug/ml overnight at 4 degree 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 1:10000 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 17KD. The expected band size for TNF alpha is at 17KD.)

Rabbit anti-Human TNF alpha Polyclonal Antibody | anti-TNF alpha antibody

Anti-TNF alpha Antibody

Gene Names
TNF; DIF; TNFA; TNFSF2; TNLG1F; TNF-alpha
Reactivity
Human
Applications
Western Blot, ELISA
Purity
Immunogen Affinity Purified
Synonyms
TNF alpha; Polyclonal Antibody; Anti-TNF alpha Antibody; Tumor necrosis factor; Cachectin; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; membrane form; N-terminal fragment; NTF; Intracellular domain 1; ICD1; Intracellular domain 2; ICD2; C-domain 1; C-domain 2; soluble form; TNF; TNFA; TNFSF2; anti-TNF alpha antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
IgG
Specificity
No cross reactivity with other proteins.
Purity/Purification
Immunogen Affinity Purified
Form/Format
Lyophilized
Sequence Length
233
Applicable Applications for anti-TNF alpha antibody
Western Blot (WB), ELISA Capture (EIA)
Application Notes
WB: 0.1-0.5 mug/ml
ELISA (Cap):1-5 mug/ml
Tested Species: In-house tested species with positive results.
Immunogen
E Coli-derived human TNF alpha recombinant protein (Position: V77-L233).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Relevant Detection Systems
It is recommended to use an Enhanced Chemiluminescent Kit with anti-Rabbit IgG (MBS176460) for Western blot.
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time.
Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. 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.Lane 1: recombinant human TNF alpha protein 1ng.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 ug/ml overnight at 4 degree 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 1:10000 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 17KD. The expected band size for TNF alpha is at 17KD.)

Western Blot (WB) (Figure 1. 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.Lane 1: recombinant human TNF alpha protein 1ng.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 ug/ml overnight at 4 degree 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 1:10000 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 17KD. The expected band size for TNF alpha is at 17KD.)
Related Product Information for anti-TNF alpha antibody
Description: Rabbit IgG polyclonal antibody for TNF alpha detection. Tested with WB, ELISA (Cap) in Human.
Background: TNF (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.
References
1. Shirai, T., Yamaguchi, H., Ito, H., Todd, C. W., Wallace, R. B.Cloning and expression in Escherichia coli of the gene for human tumour necrosis factor.Nature 313: 803-806, 1985. 2. Pennica, D., Nedwin, G. E., Hayflick, J. S., Seeburg, P. H., Derynck, R., Palladino, M. A., Kohr, W. J., Aggarwal, B. B., Goeddel, D. V.Human tumour necrosis factor: precursor structure, expression and homology to lymphotoxin.Nature 312: 724-729, 1984.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
25,644 Da
NCBI Official Full Name
tumor necrosis factor
NCBI Official Synonym Full Names
tumor necrosis factor
NCBI Official Symbol
TNF
NCBI Official Synonym Symbols
DIF; TNFA; TNFSF2; TNLG1F; TNF-alpha
NCBI Protein Information
tumor necrosis factor
UniProt Protein Name
Tumor necrosis factor
UniProt Gene Name
TNF
UniProt Synonym Gene Names
TNFA; TNFSF2; TNF-a; NTF; ICD1; ICD2

NCBI Description

This 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. [provided by RefSeq, Jul 2008]

Uniprot Description

Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Upregulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:22517918, PubMed:16829952, PubMed:23396208). Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance ().

Research Articles on TNF alpha

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Product Notes

The TNF alpha tnf (Catalog #AAA1751197) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-TNF alpha Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's TNF alpha can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), ELISA Capture (EIA). WB: 0.1-0.5 mug/ml ELISA (Cap):1-5 mug/ml Tested Species: In-house tested species with positive results. Researchers should empirically determine the suitability of the TNF alpha tnf for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TNF alpha, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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