Product nameAnti-TNF alpha antibody
See all TNF alpha primary antibodies
DescriptionRabbit polyclonal to TNF alpha
Tested applicationsSuitable for: IHC-Fr, IHC-P, WB, Neutralising, Sandwich ELISA, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment corresponding to Mouse TNF alpha aa 80-235. E.coli-derived recombinant MurineTNF-alpha
Database link: P06804
- Recombinant mouse TNF alpha protein (ab9740) can be used as a positive control in WB. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: A431
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 instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Storage bufferPBS, pH 7.4, no preservative, sterile filtered
Concentration information loading...
PurityImmunogen affinity purified
Light chain typeunknown
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.
|IHC-Fr||Use at an assay dependent concentration. PubMed: 18458097|
|IHC-P||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.
To detect TNF 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 TNF 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.
|Neutralising||Use at an assay dependent concentration. 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.|
|Sandwich ELISA||Use at an assay dependent concentration.
To detect Murine TNF alpha by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of ab9739 is required. This antigen affinity purified antibody, in conjunction with a suitable detection antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant Murine TNF alpha.
|ICC/IF||Use a concentration of 5 µg/ml.|
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.
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.
- Information by UniProt
- APC1 antibody
- APC1 protein antibody
- Cachectin antibody
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
All lanes : 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?
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.
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.
Immunohistochemical analysis of colchicine injected mouse brain (hippocampus CA1 region) tissue using ab9739 at 1.0 μg/ml overnight at 4˚C. This was followed by a peroxidase conjugated secondary antibody and then a fluorescein Tyramide Signal Amplification (TSA™) reagent. Optimal concentrations and conditions may vary.
This product has been referenced in:
- Zou W et al. Blocking meningeal lymphatic drainage aggravates Parkinson's disease-like pathology in mice overexpressing mutated a-synuclein. Transl Neurodegener 8:7 (2019). Read more (PubMed: 30867902) »
- Mann V et al. Changes in Human Foetal Osteoblasts Exposed to the Random Positioning Machine and Bone Construct Tissue Engineering. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30889841) »