Anti-TNF alpha antibody [2C8] (ab8348)

Overview

• Product nameAnti-TNF alpha antibody [2C8]See all TNF alpha primary antibodies ...
• Description
  Mouse monoclonal [2C8] to TNF alpha
• SpecificityThis antibody recognises human TNF-alpha
• Tested applicationsFlow Cyt, Neutralising, ELISA, IHC-P, IHC-Fr, ICC/IF, Sandwich ELISA more details
• Species reactivity
  Reacts with: Human
  
• Immunogen

  Human recombinant tumor necrosis factor of alpha type

• Positive controlThis antibody gave a positive result when used in the following formaldehyde fixed cell lines: A431

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPBS with 0.1% sodium azide, pH 7.4
• Concentration information loading...
• PurityProtein A purified
• Clonality Monoclonal
• Clone number2C8
• Myelomax63-Ag8.653
• IsotypeIgG1
• Light chain typeunknown
• Research Areas
  • Metabolism
  • Types of disease
  • Metabolic disorders

  • Cancer
  • Growth factors
  • TNF

  • Signal Transduction
  • Growth Factors/Hormones
  • TNF

  • Cardiovascular
  • Atherosclerosis
  • Vascular Inflammation
  • Inflammatory mediators

  • Immunology
  • Innate Immunity
  • Cytokines
  • TNF Superfamily

Applications

Target

• FunctionCytokine 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.
• Involvement in diseaseGenetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).
• Sequence similaritiesBelongs to the tumor necrosis factor family.
• Post-translational
  modificationsThe soluble form derives from the membrane form by proteolytic processing.
  The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
  O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.
• Cellular localizationSecreted and Cell membrane.

Target information above from: UniProt accession P01375 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 7124 Human
  • Omim: 191160 Human
  • SwissProt: P01375 Human
  • Unigene: 241570 Human

• Alternative names
  APC1 antibodyAPC1 protein antibodyCachectin antibody

  DIF antibodyDifferentiation inducing factor antibodyMacrophage cytotoxic factor antibodyMCF antibodyNecrosin antibodyTNF a antibodyTNF alpha antibodyTNF antibodyTNF Macrophage Derived antibodyTNF Monocyte Derived antibodyTNF Superfamily Member 2 antibodyTNF superfamily, member 2 antibodyTNF, macrophage derived antibodyTNF, monocyte derived antibodyTNF-a antibodyTNF-alpha antibodyTNFA antibodyTNFA_HUMAN antibodyTNFSF2 antibodyTumor necrosis factor alpha antibodyTumor necrosis factor antibodyTumor necrosis factor ligand superfamily member 2 antibodyTumor Necrosis Factor Precursor antibodyTumor necrosis factor, soluble form antibodyTumour Necrosis Factor Alpha antibody

  see all

Anti-TNF alpha antibody [2C8] images

• 

  Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [2C8] (ab8348)

  ICC/IF image of ab8348 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab8348 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• 

  Sandwich ELISA - TNF alpha antibody [2C8] (ab8348)
  Standard curve for TNF alpha (Analyte: ab9642); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [2C8] to TNF alpha (ab8348) at 5µg/ml and Detector Antibody Rabbit polyclonal to TNF alpha (ab9635) at 0.5µg/ml.
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - TNF alpha antibody [2C8] (ab8348)
  Ab8348 staining human tonsil. Staining is localized to the plasma membrane and cytoplasm.
  Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
  Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
• 

  Flow Cytometry - Anti-TNF alpha antibody [2C8] (ab8348)
  Overlay histogram showing THP1 cells stained with ab8348 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8348, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  
  

Protocols

Protocol Booklet