Anti-TNF alpha antibody (ab6671)

Rabbit polyclonal TNF alpha antibody. Validated in WB, ELISA, IHC, Neut, ICC/IF and tested in Mouse, Rat, Guinea pig, Human, Pig, Fish, Cynomolgus monkey. Cited in 230 publication(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to TNF alpha
  • Host species
    Rabbit
  • Specificity

    The immunogen used to raise this antibody shares 78% homology with mouse and rat TNFalpha protein. Some customers have successfully used ab6761 with mouse and rat samples, however we do not batch test in these species and therefore we cannot guarantee this product will consistently work with mouse and rat.  We would recommend ab205587 or ab1793 as an alternative product for use with these species.

  • Tested applications
    Suitable for: ELISA, IHC-P, IHC-Fr, Neutralising, WB, ICC/IF, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Human, Cynomolgus monkey
    Predicted to work with: Mouse, Rat, Dog, Pig, Monkey, Non human primates
  • Immunogen

    Recombinant full length protein corresponding to TNF alpha.
    Database link: P01375

  • Positive control
  • General notes
    Endotoxin content by LAL is <10 pg/ml.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab6671 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
ELISA 1/200 - 1/1000.
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/100 - 1/200.
Neutralising 1/200.
WB 1/500 - 1/2000.

Membrane Blocking is recommended with BSA not Milk for this product.

Suitable for use as a positive control for Western blot against recombinant TNFalpha produced in E.Coli. 

ICC/IF Use at an assay dependent concentration. PubMed: 19458984
IHC-FoFr Use at an assay dependent concentration.

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

  • ab6671 staining TNF alpha in Human testis & colon CA tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% H2O2 for 10 minutes at 25°C; antigen retrieval was by heat mediation using EDTA, pH9, 100C, 20 min. Samples were incubated with primary antibody (1/75) for 20 minutes at 25°C. An undiluted HRP polymer-conjugated Goat anti-mouse anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • ab6671 staining TNF alpha in Guinea pig Lung and heart cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formalin and blocked with 1% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA in PBST) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated Donkey anti-rabbit IgG (H+L) polyclonal (1/800) was used as the secondary antibody.

    See Abreview

  • ab6671 staining TNF alpha in Cynomolgus Monkey dendritic cells/macrophages from inlamed skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 20 minutes at room temperature; antigen retrieval was by heat mediation in citrate buffer, pH6.0. Samples were incubated with primary antibody (1/100) for 30 minutes at room temperature. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/2000) was used as the secondary antibody.

    See Abreview

  • Anti-TNF alpha antibody (ab6671) at 1/1000 dilution + Recombinant TNF-alpha protein

    Secondary
    Goat Anti-Rabbit Dylight® 649 Conjugate at 1/20000 dilution

    Additional bands at: 16 kDa (possible cleavage fragment)



    Western Blot analysis, labelling TNF-alpha with ab6671 at 1/1000. Blocking was with 1% BSA in TBS-T incubated for 30 minutes at room temperature. The Cleavage Fragment is related to the soluble isoform 77 - 223. 

  • ab6671 staining human artery tissue sections by IHC-P.  Sections were fixed in formaldehyde and subjected to heat mediated antigen retrieval in citrate buffer (pH 6.0) prior to blocking with 1.5% serum for 10 minutes.  The primary antibody was diluted 1/100 and incubated with the sample for 24 hours at 4°C.  An HRP-conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • ab6671 staining TNF alpha from human colon by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Cells were formaldehyde fixed prior to blocking in 10% serum for 2 hours at 21°C. The primary antibody was diluted 1/100 and incubated with the sample for 2 hours at 21°C. Alexa fluor® 680 goat polyclonal, diluted 1/5000, was used as the secondary.

    See Abreview

  • Immunohistochemical analysis of formalin-fixed, paraffin embedded fish tissue, staining TNF alpha with ab6671.

    Tissue was fixed with Bouin's fixative; antigen retrieval was by heat mediation in a citrate buffer (pH 6). Samples were incubated with primary antibody (1/600 in diluent) for 2 hours. An HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Dormer NH  et al. Evaluation of a transtympanic delivery system in Mus musculus for extended release steroids. Eur J Pharm Sci 126:3-10 (2019). Read more (PubMed: 29329746) »
  • Sayyaf Dezfuli B  et al. Description of epithelial granular cell in catshark spiral intestine: Immunohistochemistry and ultrastructure. J Morphol 280:205-213 (2019). Read more (PubMed: 30589112) »
See all 289 Publications for this product

Customer reviews and Q&As

1-10 of 55 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric
Permeabilization
No
Specification
colon
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 17 2019

Application
Western blot
Sample
Mouse Tissue lysate - whole (Lung)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
50 µg
Specification
Lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 07 2019

Abreviews
Application
Western blot
Sample
Goat Tissue lysate - other (Goat MSCs and Goat Nucleus pulposus cells pellets)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
20 µg
Treatment
1 ug/mL Lipopolysaccharide induced 3 days
Specification
Goat MSCs and Goat Nucleus pulposus cells pellets
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 26°C

Dr. Chenghao Zhang

Verified customer

Submitted Sep 07 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (OV90 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)
Loading amount
50 µg
Specification
OV90 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 18 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (vessel)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: H2O2
Permeabilization
No
Specification
vessel
Blocking step
dako antibody diluent with background reducing components as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 30 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Neuroblastoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 8.5
Permeabilization
No
Specification
Neuroblastoma
Fixative
Formaldehyde

Mr. Allan Wang

Verified customer

Submitted Apr 18 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (molar injury)
Permeabilization
Yes - Triton X
Specification
molar injury
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Tina Yuan

Verified customer

Submitted Jan 25 2018

Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (brain)
Specification
brain

Abcam user community

Verified customer

Submitted Apr 24 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Human Synoviocyte fibroblast-like cell)
Gel Running Conditions
Reduced Denaturing (9% SDS-PAGE)
Loading amount
20 µg
Treatment
D-glucose 500 mg/dl for 24hrs
Specification
Human Synoviocyte fibroblast-like cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 27 2016

Application
Western blot
Sample
Chicken Tissue lysate - whole (thymus)
Gel Running Conditions
Reduced Denaturing (10-20% Tris-glycine)
Loading amount
20 µg
Specification
thymus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Jenny Shoots

Verified customer

Submitted Dec 07 2015

1-10 of 55 Abreviews or Q&A

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