Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21753-109] to TNF alpha
- Suitable for: ICC/IF, WB, Flow Cyt, IP
- Reacts with: Mouse, Rat
Product nameAnti-TNF alpha antibody [EPR21753-109]
See all TNF alpha primary antibodies
DescriptionRabbit monoclonal [EPR21753-109] to TNF alpha
The protein level of TNF alpha in normal samples is very weak. The TNF alpha expression must be stimulated.
Tested applicationsSuitable for: ICC/IF, WB, Flow Cyt, IPmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Mouse, Rat
Recombinant fragment within Rat TNF alpha aa 50 to the C-terminus. The exact sequence is proprietary.
Database link: P16599
- WB: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate; RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h, whole cell lysate. ICC/IF: RAW 264.7 cells. Flow: RAW 264.7 cells. IP: Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h, whole cell lysate
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab205587 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Predicted molecular weight: 26 kDa.|
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
TNF alpha was immunoprecipitated from 0.35 mg rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate with ab205587 at 1/50 dilution. Western blot was performed on the immunoprecipitate using ab205587 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
Lane 1: Rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate 10μg (input).
Lane 2: ab205587 IP in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205587 in rat splenocytes (treated with 20ng/ml PMA, 1μg/ml Ionomycin and 10μM BFA for 6h) whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
The MW observed is consistent with what has been described in the literature (PMID: 9933416).
Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h (Red) / Untreated control (Green), compared to an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue) and an isotype control - Rabbit monoclonal IgG (ab172730, Black). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077)at 1/2000 dilution was used as the secondary antibody.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (+/- treatment with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h) cells labeling TNF alpha with ab205587 at 1/100 dilution, followed by a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cells treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h. The Nuclear counterstain is DAPI (Blue). Tubulin was stained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
All lanes : Anti-TNF alpha antibody [EPR21753-109] (ab205587) at 1/1000 dilution
Lane 1 : Untreated rat splenocytes whole cell lysate
Lane 2 : Rat splenocytes treated with 20ng/ml PMA, 1µg/ml Ionomycin and 10µM BFA for 6h whole cell lysate
Lane 3 : Untreated RAW 264.7(mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 4 : RAW 264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 26 kDa
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 20 seconds
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 9933416)
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab205587 has not yet been referenced specifically in any publications.