Overview

  • Product nameAnti-TNF alpha antibodySee all TNF alpha primary antibodies ...
  • Description
    Rabbit polyclonal to TNF alpha
  • Tested applicationsIHC-Fr, IHC-P, WB, ELISA, Neutralising, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Highly pure (>98%) recombinant mTNF-alpha (mouse Tumor Necrosis Factor-alpha).

  • Positive control
    • This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: A431

Properties

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 Abreviews Notes
IHC-Fr Use at an assay dependent dilution. PubMed: 18458097
IHC-P Use at an assay dependent dilution.
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 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 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 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
    modifications
    The 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
  • Database links
  • Alternative names
    • APC1 antibody
    • APC1 protein antibody
    • Cachectin antibody
    • DIF antibody
    • Differentiation inducing factor antibody
    • Macrophage cytotoxic factor antibody
    • MCF antibody
    • Necrosin antibody
    • TNF a antibody
    • TNF alpha antibody
    • TNF antibody
    • TNF Macrophage Derived antibody
    • TNF Monocyte Derived antibody
    • TNF Superfamily Member 2 antibody
    • TNF superfamily, member 2 antibody
    • TNF, macrophage derived antibody
    • TNF, monocyte derived antibody
    • TNF-a antibody
    • TNF-alpha antibody
    • TNFA antibody
    • TNFA_HUMAN antibody
    • TNFSF2 antibody
    • Tumor necrosis factor alpha antibody
    • Tumor necrosis factor antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor Necrosis Factor Precursor antibody
    • Tumor Necrosis Factor, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    • Tumour Necrosis Factor Alpha antibody
    see all

Anti-TNF alpha antibody images

  • 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

    See Abreview

  • 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.

References for Anti-TNF alpha antibody (ab9739)

This product has been referenced in:
  • Chen S  et al. SOX2 regulates apoptosis through MAP4K4-Survivin signaling pathway in human lung cancer cells. Carcinogenesis 35:613-23 (2014). WB ; Human . Read more (PubMed: 24233838) »
  • Schmidt N  et al. Dietary vitamin D inadequacy accelerates calcification and osteoblast-like cell formation in the vascular system of LDL receptor knockout and wild-type mice. J Nutr 144:638-46 (2014). Mouse . Read more (PubMed: 24647396) »

See all 13 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Non-Denaturing (Native)
Sample Mouse Serum (Mouse Serum)
Specification Mouse Serum
Blocking step Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Mr. FRANCESCO ELIA MARINO

Verified customer

Submitted Jul 30 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Jul 08 2014

Thank you for contacting us.

Unfortunately, we cannot test every organ or tissue when we establish a product.However, if the knee joints were decalcified properly and the species is tested, I don't see any problems why these antibodies shoul...

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All of these antibodies recognise cytokines which are present in inflammed tissue therefore any inflammed tissue would be a good positive control.

In our catalogue, we have several cancer slides which would be appropriate:
tonsil: (ab514...

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Vielen Dank für Ihre Anfrage.

Ich habe vier primäre AK für Sie herausgesucht, die alle in einer relativ hohen Verdünnung eingesetzt werden können, so dass sie für eine Weile reichen sollten. Das Sie alle Proben nu...

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Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unserem Wissen nach wurde ab17442 bisher noch nicht in sandwichELISA getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zurzeit ein spezielles...

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Thank you for contacting us and for your interest in our products.

As far as we are aware, ab9739 has never been tested in IP. All tested applications covered by the 6 month guarantee are specified on our datasheets, and these are updated as...

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Thank you very much for your reply.

You are correct, and I apologize for this mistake. I converted the 100 micrograms into milligrams, not grams as it should have been for the calculation. There should be 4.01e14 molecules per vial. Thank you...

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Thank you very much for your reply.

There are approximately 4.01 x 10^17 molecules of anti-TNF alpha IgG per 100microgram vial. This is calculated as:

Molecular weight of IgG= 150 kDa= 150,000 g/mol

100 micrograms divided ...

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Thank you very much for your call and for your enquiry.

Ab9739 was raised against the recombinant soluble form of murine TNF alpha. As this is a polyclonal antibody and the soluble form shares homology with the membrane form, it should bind t...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"