Recombinant
RabMAb

Recombinant Anti-TNF alpha antibody [EPR21753-109] (ab205587)

Overview

  • Product name

    Anti-TNF alpha antibody [EPR21753-109]
    See all TNF alpha primary antibodies
  • Description

    Rabbit monoclonal [EPR21753-109] to TNF alpha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Rat TNF alpha aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: P16599

  • Positive control

    • 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
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
ICC/IF 1/100.
WB 1/1000. Predicted molecular weight: 26 kDa.
Flow Cyt 1/100.
IP 1/50.
  • Application notes
    Is unsuitable for IHC-P.
  • 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 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 (TNF superfamily member 2) 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

    • 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.

      Secondary
      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)

    References

    ab205587 has not yet been referenced specifically in any publications.

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