Overview

  • Product name

    Anti-MCP1 antibody [EPR21025]
    See all MCP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21025] to MCP1
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MCP1 aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: P13500

  • Positive control

    • WB: THP-1 treated with PMA (ab120297), LPS, and BFA whole cell lysate. ICC/IF: THP-1 cells treated with PMA (ab120297), LPS, and BFA. Flow Cyt: THP-1 treated with PMA (ab120297), LPS, and BFA. IP: THP-1 treated with PMA (ab120297), LPS, and BFA 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 ab214819 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
WB 1/1000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
ICC/IF 1/50.
Flow Cyt 1/500.
IP 1/30.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      Chemotactic factor that attracts monocytes and basophils but not neutrophils or eosinophils. Augments monocyte anti-tumor activity. Has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis or atherosclerosis. May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.
    • Sequence similarities

      Belongs to the intercrine beta (chemokine CC) family.
    • Post-translational
      modifications

      Processing at the N-terminus can regulate receptor and target cell selectivity. Deletion of the N-terminal residue converts it from an activator of basophil to an eosinophil chemoattractant.
    • Cellular localization

      Secreted.
    • Information by UniProt
    • Database links

    • Alternative names

      • C-C motif chemokine 2 antibody
      • CCL2 antibody
      • CCL2_HUMAN antibody
      • Chemokine (C C motif) ligand 2 antibody
      • GDCF 2 antibody
      • GDCF-2 antibody
      • GDCF2 antibody
      • HC11 antibody
      • HSMCR30 antibody
      • JE antibody
      • MCAF antibody
      • MCP 1 antibody
      • MCP-1 antibody
      • MCP1 antibody
      • MGC9434 antibody
      • Monocyte chemoattractant protein 1 antibody
      • Monocyte chemotactic and activating factor antibody
      • Monocyte chemotactic protein 1 antibody
      • Monocyte secretory protein JE antibody
      • SCYA2 antibody
      • Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig je) antibody
      • Small inducible cytokine A2 antibody
      • Small inducible cytokine subfamily A (Cys Cys), member 2 antibody
      • Small-inducible cytokine A2 antibody
      • SMC CF antibody
      • SMC-CF antibody
      • SMCCF antibody
      see all

    Images

    • All lanes : Anti-MCP1 antibody [EPR21025] (ab214819) at 1/1000 dilution

      Lane 1 : Untreated THP-1 (human monocytic leukemia cell line) whole cell lysate
      Lane 2 : THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours, then treated with 100ng/ml lipopolysaccharide (LPS) for 7 hours, then with 1 µg/ml Brefeldin A (BFA) added after 4 hours, 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: 11 kDa
      Observed band size: 11 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia cell line) cells, untreated or treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24 hours, then treated with 100ng/ml lipopolysaccharide (LPS) for 7 hours, with 1 μg/ml Brefeldin A (BFA) added after 4 hours, labeling MCP1 with ab214819 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in THP-1 treated cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • MCP1 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24h, then treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1μg/ml Brefeldin A (BFA) for another 3h whole cell lysate with ab214819 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214819 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution

      Lane 1: THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24h, then treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1μg/ml Brefeldin A (BFA) for another 3h whole cell lysate 10 µg (Input). 

      Lane 2: ab214819 IP in THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24h, then treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1μg/ml Brefeldin A (BFA) for another 3h whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214819 in THP-1 treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24h, then treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1μg/ml Brefeldin A (BFA) for another 3h whole cell lysate.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20-permeabilized THP-1 (human monocytic leukemia cell line) cell line, treated with 80nM Phorbol-12-myristate-13-acetate (PMA, ab120297) for 24h, then treated with 100ng/ml lipopolysaccharide (LPS) for 4h, then together with 1μg/ml Brefeldin A (BFA) for another 3h (Right) / Untreated control (Left) labeling MCP1 with ab214891 at 1/500 dilution. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    References

    ab214819 has not yet been referenced specifically in any publications.

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