Overview
- Product nameAnti-TNF alpha antibodySee all TNF alpha primary antibodies ...
- DescriptionRabbit polyclonal to TNF alpha
- Tested applicationsIHC-Fr, IHC-P, WB, ELISA, Neutralising, ICC/IF more details
- Species reactivityReacts with: Mouse, Human
- Immunogen
Highly pure (>98%) recombinant mTNF-alpha (mouse Tumor Necrosis Factor-alpha).
- Positive controlThis antibody gave a positive result in IF in the following Formaldehyde fixed cell line: A431
Properties
- FormLyophilised:Reconstitute with 200µl of sterile water. Please note that if you receive this product in liquid form it has already been reconstituted as described and no further reconstitution is necessary.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
- Storage bufferPBS, pH 7.4, no preservative, sterile filtered
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Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- Isotypeunknown
- Light chain typeunknown
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab9739 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| IHC-Fr | IHC-Fr: Use at an assay dependent dilution. PubMed: 18458097 |
| IHC-P | IHC-P: Use at an assay dependent dilution. |
| WB | WB: Use at an assay dependent dilution. To detect mTNF-alpha by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant mTNF-alpha is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions. The presursor is ~26 kDa and the secreted form is ~17 kDa. |
| ELISA | ELISA: Use at an assay dependent dilution. To detect mTNF-alpha by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant mTNF-alpha. |
| Neutralising | Neut: Use at an assay dependent dilution. To yield one-half maximal inhibition [ND50] of the biological activity of mTNF-alpha (0.25 ng/ml), a concentration of 0.08 – 0.10 µg/ml of this antibody is required. |
| ICC/IF | ICC/IF: Use a concentration of 5 µg/ml. |
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.
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Database links
- Entrez Gene: 7124 Human
- Entrez Gene: 21926 Mouse
- Omim: 191160 Human
- SwissProt: P01375 Human
- SwissProt: P06804 Mouse
- Unigene: 241570 Human
- Unigene: 1293 Mouse
Target information above from: UniProt accession
P01375
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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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 images
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Western blot - TNF alpha antibody (ab9739)All lanes : Anti-TNF alpha antibody (ab9739) at 1/3500 dilution
Lane 1 : 100ug LPS stimulated J774.A1 mice macrophages
Lane 2 : 50ug LPS stimulated J774.A1 mice macrophages
Lane 3 : 25ug LPS stimulated J774.A1 mice macrophages
Secondary
HRP conjugated goat anti-rabbit antibody
Performed under reducing conditions.
Observed band size : 17 kDa (why is the actual band size different from the predicted?)This image is courtesy of an Abreview submitted by Dr sandra sobocanec
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-TNF alpha antibody(ab9739)ab9739 (2µg/ml) staining TNF alpha in human tonsil using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining within the germinal follicle.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody (ab9739)ICC/IF image of ab9739 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 ab9739 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.
References for Anti-TNF alpha antibody (ab9739)
This product has been referenced in:
- Altintas MM et al. Mast cells, macrophages, and crown-like structures distinguish subcutaneous from visceral fat in mice. J Lipid Res 52:480-8 (2011). Read more (PubMed: 21148461) »
- Yang B et al. Sh3pxd2b mice are a model for craniofacial dysmorphology and otitis media. PLoS One 6:e22622 (2011). IHC-P ; Mouse . Read more (PubMed: 21818352) »
