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

1-10 of 16 Abreviews or Q&A

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

Answer

Thank you for contacting Abcam.

The antibody is covered under our Abpromise for six months and is guaranteed to work in IHC-P on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

To be able to help you further with this matter and to see if there is any protocol advice I could give to resolve the issue, would you be able to answer a few more protocol questions and also specify the species that your samples came from:

1) Fixation step

Fixation time

Fixation temperature

2) Antigen retrieval method (time and procedure)

3) Permeabilization method:

Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

Permeabilizing agent and concentration:

4) Blocking agent (eg BSA, serum…):

Concentration

Blocking time

Blocking temperature

5) Endogenous peroxidases blocked?

Endogenous biotins blocked?

6) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution

Diluent buffer

Incubation time

7) Secondary antibody:

Species:

Reacts against:

Concentration or dilution

Diluent buffer

Incubation time

Fluorochrome or enzyme conjugate

8) Washing after primary and secondary antibodies:

Buffer

Number of washes

9) Detection method

10) How many times have you run this staining?

Do you obtain the same results every time?

What steps have you altered to try and optimize the use of this antibody?

As mentioned previously, if we cannot resolve the issues that you are having with ab9669, and you are working in samples that we guarantee, then we will be happy to either replace or refund the cost of the antibody.

Read More

Answer

Thank you for your reply, and I apologize for this inconvenience.

I certainly understand that ab9858 is not as useful to you as ab73680 would be. I can issue a credit to you.

The vial of ab8020 should be arriving shortly, please keep me updated about the results with this replacement, and let me know if there is anything else that you need!

Read More

Answer

Thank you again for your patience.

I apologize, but ab73680 is out of stock, and it may not be put back into stock for several months as recent lots have not passed quality control testing. Would you be interested in trying another vial of ab9669, or ab9858-

https://www.abcam.com/MCP1-antibody-ab9858.html

Please let me know and I will arrange for this to be sent to you promptly. Have a great day!

Read More

Answer

Thank you for your reply.

Please keep me updated on any progress. I hope that we hear something from them soon.

Please let me know if you need anything else from me at this time, and I'll let you know if I hear from someone at BioConnect. You have a good weekend as well!

Read More

Answer

Thank you again for your patience.

At this point, I have not heard from Bio-Connect regarding these issues and I've looked through our system and don't see that they've corresponded with anyone else at Abcam. Have you received any update from them? Is there anyone at Bio-Connect who I can contact?

I look forward to hearing from you. I'll be out of the office Monday-Thursday of next week but I will be back in touch nextFriday if there is anything else that I can do to help resolve these issues. Thanks so much! Have a great weekend.

Read More

Answer

Thank you for getting back to me and for getting in touch with BioConnect. I will let you know if I don't hear anything from them shortly, and we will figure out the vial of ab9669. I'll also send you a confirmation email after I work everything out with them.

Thank you for your understanding, and let me know if you have any questions in the meantime.

Read More

Answer

Thank you for your reply and for confirming that information.

I'm processing the refunds and replacements now. I think I've found your order for ab22048, ab9100, and ab8021, but I am having some trouble finding the order for ab9669. Were these ordered through our distributor in the Netherlands, Bio-Connect? If so, could you plrease forward me your shipping address and your Bio-Connect customer or account number, to make sure the refunds and replacements reach you.

For the MCP1 antibody, is it possible that you currently have a different antibody other than ab9669? Our records only show one order of ab9669 shipped to the Netherlands and that was placed in 2008. If you could find out the Abcam order number from your contact at Bio-Connect, it would be very helpful.

I am sorry for the delay in processing these, but I just want to make sure you are properly reimbursed. I look forward to hearing from you, and please let me know if you have any questions or further information. I hope you enjoy your weekend as well!

Read More

Answer

Thank you for getting back to me with your selections.

For CX3CR1, we do have a couple of other anti-human antibodies that have been tested in IHC on FFPE sections, ab8020 and ab51668-

https://www.abcam.com/CX3CR1-antibody-ab8020.html

https://www.abcam.com/CX3CR1-antibody-ab51668.html

Would you be interested in trying one of these as a replacement for ab8021? If not, I will go ahead and proceed with the refund. I will process your requests as soon as I hear back from you!

I look forward to hearing from you. Please let me know if you need anything else, and have a great day!

Read More

1-10 of 16 Abreviews or Q&A

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

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