Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab9669 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 Use a concentration of 0.1 - 0.2 µg/ml.

To detect hMCAF/MCP-1 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 MCP-1 is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.

Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal to MCP1 (ab9858).

To detect MCP-1 by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of ab9669 is required. This antigen affinity purified antibody, in conjunction with ab9858 as a capture antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant MCP-1.

IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 20369226

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

  • HL-60 cells were incubated at 37°C for 24hrs with vehicle control (0 µM) and varied concentrations of 2-Arachidonylglycerol (ab120098). Increased expression of MCP1 in HL-60 cells correlates with an increase in 2-Arachidonylglycerol concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab9669 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

  • ab9669 at 2.5 µg/ml staining human breast invasive ductal carcinoma tissue by IHC-P.  The anitbody was incubated for two hours at room temperature. An HRP-labeled polymer detection system was used with a DAB chromogen. Heat induced antigen retrieval with a pH 6.0 Sodium Citrate buffer is recommended. 
  • Standard curve for MCP1 (Analyte: ab73866); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal to MCP1 (ab9858) at 1µg/ml and Detector Antibody Rabbit polyclonal to MCP1 (ab9669) at 0.5µg/ml.
  • All lanes : Anti-MCP1 antibody (ab9669) at 0.2 µg/ml

    Lane 1 : Recombinant human MCP1 protein (ab73866) at 0.01 µg
    Lane 2 : Recombinant human MCP1 protein (ab73866) at 0.001 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 1 minute
  • Anti-MCP1 antibody (ab9669) at 1/4000 dilution + lysate prepared from human smooth muscle treated with 100µg/ml glycated LDL at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/20000 dilution

    Developed using the ECL technique.

    Observed band size: 12-14 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute

    See Abreview

References

This product has been referenced in:
  • Wei CC  et al. Allergic Conjunctivitis-induced Retinal Inflammation Promotes Myopia Progression. EBioMedicine 28:274-286 (2018). Read more (PubMed: 29398596) »
  • Weiler J  et al. Matrix metalloproteinase-9 (MMP9) is involved in the TNF-a-induced fusion of human M13SV1-Cre breast epithelial cells and human MDA-MB-435-pFDR1 cancer cells. Cell Commun Signal 16:14 (2018). WB . Read more (PubMed: 29636110) »
See all 33 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 25% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 18 2017

Application
Western blot
Sample
Human Cell lysate - other (Endothelial cells (EA.hy926) - conditioned media)
Gel Running Conditions
Reduced Denaturing (12% SDS-PAGE)
Loading amount
500000 cells
Treatment
100 µg/ml glycated LDL (AGE-LDL) for 24 hours
Specification
Endothelial cells (EA.hy926) - conditioned media
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Anca Sima

Verified customer

Submitted Jun 18 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (smooth muscle cell-conditioned medium)
Loading amount
10 µg
Specification
smooth muscle cell-conditioned medium
Treatment
100 ug/ml glycated LDL
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Laura Toma

Verified customer

Submitted Feb 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Fetal foreskin cells)
Loading amount
10 µg
Specification
Fetal foreskin cells
Gel Running Conditions
Reduced Denaturing (4-20% gradient)
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 Feb 17 2011

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

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