Overview

  • Product name
  • Description
    Rabbit polyclonal to TNF alpha
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, WB, Neutralising, Sandwich ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to Mouse TNF alpha aa 80-235. E.coli-derived recombinant MurineTNF-alpha
    Database link: P06804

  • 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 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.

Target

  • Function
    Cytokine 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 disease
    Genetic 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 similarities
    Belongs 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 localization
    Secreted 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
    • Tnf 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, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    see all

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.

  • 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.

References

This product has been referenced in:
  • Freitas LS  et al. In situ detection of Chlamydia pneumoniae, C. trachomatis, and cytokines among cardiovascular diseased patients from the Amazon region of Brazil. Infect Drug Resist 10:109-114 (2017). IHC-P ; Human . Read more (PubMed: 28435302) »
  • Zhang Y  et al. An Extract from Shrimp Processing By-Products Protects SH-SY5Y Cells from Neurotoxicity Induced by Aß25-35. Mar Drugs 15:N/A (2017). WB ; Human . Read more (PubMed: 28327516) »

See all 34 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse lung)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
Mouse lung after 24-hour ischemia
Specification
Mouse lung
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 Mar 03 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (breast tumor)
Permeabilization
No
Specification
breast tumor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 15 2017

Application
Western blot
Sample
Mouse Serum (Mouse Serum)
Gel Running Conditions
Reduced Non-Denaturing (Native)
Loading amount
20 µg
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

Abcam user community

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

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Recombinant protein (Recombinant Human TNF-alpha produced in E.coil)
Loading amount
0.5 µg
Specification
Recombinant Human TNF-alpha produced in E.coil
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 24 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (lungs)
Loading amount
25 µg
Specification
lungs
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (J774.A1 mice macrophages)
Loading amount
50 µg
Specification
J774.A1 mice macrophages
Treatment
stimulated for 24h with 1 µg/ml LPS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Sandra Sobocanec

Verified customer

Submitted Jan 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (skin)
Loading amount
100 µg
Specification
skin
Blocking step
Other as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 50
Username

Dr. Precedia Massey

Verified customer

Submitted Apr 26 2006

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

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