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:
  • Zhang HX  et al. Thalidomide decreases high glucose-induced extracellular matrix protein synthesis in mesangial cells via the AMPK pathway. Exp Ther Med 17:927-934 (2019). Read more (PubMed: 30651882) »
  • Li YL  et al. Prognostic significance of monocyte chemoattractant protein-1 and CC chemokine receptor 2 in diffuse large B cell lymphoma. Ann Hematol 98:413-422 (2019). Read more (PubMed: 30374624) »
See all 41 Publications for this product

Customer reviews and Q&As

1-10 of 12 Q&A

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!

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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!

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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!

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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.

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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!

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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

Answer

Thank you for getting back to me, and I am happy to help resolve this issue. I look forward to hearing from you, and please let me know if you would like any assistance selecting replacements, or if you have any other questions.

Have a great day!

Read More

Question

Dear sir, madam,

april 2011 our lab ordered the following abcam antibodies

anti-human Cx3Cr1 (ab8021)

anti-human BDNF (ab72439)

anti-human TLR4 (ab22048)

anti-human TLR2 (ab9100)

anti-human MCP-1(ab9669)

anti-human NGF (ab6198)

anti-human TGF-beta (ab66043)

Of these, BDNF, NGF, TGF seem to work fine. However, after using various human tissue imbedded paraffine sections (tonsil, skin psoriasis, lymphnode, and brain sepsis) we still get no (TLR2) and vague/unspecific (MCP1,CX3CR1, TLR4) immunoreactivity (IR).

The most concentrated dilution suggested for these antibodies was used, and antigen retrieval was performed according to protocol (for example TLR4: antigen retrieval with citrate buffer PH6).
The reason I haven't contacted you before, is because I wanted to test the markers on various human tissue which I first had to gather.

Please find images attached in a 'We Tranfer file' of TLR2, TLR4, MCP1 and Cx3Cr1 IHC of cortex sepsis, skin psoriasis, tonsil and lymphnode human paraffine embedded tissue (Images were too big to attach in this email, even when compressed).

I would like your expert opinion on these stainings.
In our opninion
TLR2 shows no immunoreactivity at all (note:TLR2 datasheet explicitly states lymphnode as tissue specificity).
TLR4 shows vessel-staining in lymphnode and cortex, and broader staining in tonsil and psoriasis. However doubtdful on the specificty.
MCP1 shows a diffuse staining pattern, which again may be unspecific?
CX3CR1 shows no immunoreactivity in sepsis cortex nor lymphnode, but some IR in tonsil in psoriasis.

Since these 4 antibodies do not perform as described on the datasheet, I wanted to ask for either a refund or a replacement of these products with Anti-TLR2 (ab24192), Anti-TLR4 (ab13556), Anti-MCP1 (ab73680) and a different batch of the same CxCr1 (ab8021).

Read More
Answer

Thank you for contacting us and letting us know about the trouble with these antibodies.

After reviewing the protocol and images, I agree that these antibodies might not be performing as expected. Our guarantee period generally extends for 6 months after purchase, but I understand that it takes time to thoroughly test so many antibodies, and so I would be happy to send the replacements that you mention or to issue refund. Would prefer a replacement or refund, and could you send me your original order number or PO number?

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

1-10 of 12 Q&A

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