Overview

  • Product name
    Anti-TNF alpha antibody [52B83]
    See all TNF alpha primary antibodies
  • Description
    Mouse monoclonal [52B83] to TNF alpha
  • Specificity
    This antibody detects both natural and recombinant TNFa. It does not cross-react with TNF beta or lymphotoxin. It reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. It does not react with receptor-bound TNF-alpha. Non-specific background staining is observed in connective tissues.
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, ICC/IF, ELISA, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Guinea pig, Human, Chimpanzee, Zebrafish, Cynomolgus monkey, Rhesus monkey, Apteronotus leptorhynchus
  • Immunogen

    Full length native protein (purified) (Human).

  • Positive control
  • General notes
    Dilution of 20-200 times is useful for IHC on paraffin embedded tissues and frozen sections; flow cytometry and Western blotting. Useful for staining classic paraffin embedded tissues without antigen retrieval.

Properties

Applications

Our Abpromise guarantee covers the use of ab1793 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
Flow Cyt 1/10.

(methanol fixed cells)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr 1/50.

Fixed in acetone for 10 minutes

ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 17 kDa.

TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha  is 17.4 kDa

A reduced sample treatment and 15% SDS-Page was used.

IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • ab1793 staining TNF alpha in mouse liver tissue sections, from mice intoxicated with carbon tetrachloride for 7 weeks, by IHC-P (Formaldehyde-fixed, Paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 1% BSA for 30 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/50 in 1% BSA) at 4°C for 12 hours. An undiluted HRP-conjugated secondary antibody was used.

    See Abreview

  • ab1793 staining TNFα in RAW 264.7 cells treated with (R,S)-rolipram (ab120029), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R,S)-rolipram, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120029 ((R,S)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Overlay histogram showing RAW 264.7 cells stained with ab1793 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1793, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in RAW 264.7 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • ab1793 staining TNF alpha in Mouse duodenum and pancreatic cancer associated stroma sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were stained by DAPI.

    See Abreview

  • ab1793 staining TNF alpha in Mouse pancreatic neoplasia tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with a MOM kit for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An undiluted Biotin-conjugated Goat anti-mouse IgG polyclonal (MOM kit) was used as the secondary antibody.

    See Abreview

  • ab1793 staining TNFα in RAW 264.7 cells treated with (R)-(-)-rolipram (ab120031), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R)-(-)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120031 ((R)-(-)--rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab1793 staining TNFα in RAW 264.7 cells treated with (S)-(+)-rolipram (ab120030), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (S)-(+)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120030 ((S)-(+)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References

This product has been referenced in:
  • Andrés-Benito P  et al. Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: implications in frontotemporal lobar degeneration. Aging (Albany NY) 9:823-851 (2017). IHC-P ; Human . Read more (PubMed: 28283675) »
  • Wu NC  et al. Intravenous superoxide dismutase as a protective agent to prevent impairment of lung function induced by high tidal volume ventilation. BMC Pulm Med 17:105 (2017). Read more (PubMed: 28747201) »

See all 59 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jan 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (mcf-7)
Loading amount
1e+006 cells
Specification
mcf-7
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted May 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Apteronotus leptorhynchus Tissue sections (Spinal cord)
Specification
Spinal cord
Fixative
Paraformaldehyde
Blocking step
3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Nov 14 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary astroglial culture)
Specification
Primary astroglial culture
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Sep 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (pancreatic cancer tissue samples)
Loading amount
30 µg
Specification
pancreatic cancer tissue samples
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 Jul 22 2010

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

Abcam user community

Verified customer

Submitted Jul 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (10 embryos per well)
Loading amount
10 cells
Specification
10 embryos per well
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (pancreatic cancer samples)
Loading amount
35 µg
Specification
pancreatic cancer samples
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 Jul 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mouse pancreatic neoplasia)
Specification
mouse pancreatic neoplasia
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
No
Blocking step
No blocking step used for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Duodenum and pancreatic cancer associated stroma)
Specification
Duodenum and pancreatic cancer associated stroma
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 09 2010

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