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    mcp1-antibody-ab9669.pdf

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Immunology Innate Immunity Chemokines Alpha Chemokines (CXC)
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Anti-MCP1 antibody (ab9669)

  • Datasheet
  • SDS
Reviews (4)Q&A (12)References (65)

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Western blot - Anti-MCP1 antibody (ab9669)
  • Sandwich ELISA - Anti-MCP1 antibody (ab9669)
  • Western blot - Anti-MCP1 antibody (ab9669)
  • Western blot - Anti-MCP1 antibody (ab9669)

Key features and details

  • Rabbit polyclonal to MCP1
  • Suitable for: WB, Sandwich ELISA
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

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Overview

  • Product name

    Anti-MCP1 antibody
    See all MCP1 primary antibodies
  • Description

    Rabbit polyclonal to MCP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Sandwich ELISAmore details
  • Species reactivity

    Reacts with: Human, Recombinant fragment
  • Immunogen

    Recombinant fragment corresponding to Human MCP1.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • Purchase matching WB positive control:Recombinant human MCP1 protein
  • General notes

    Although some customers have been successful in IHC we no longer batch test in this application. For and IHC validated antibody please see ab73680

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Lyophilized:Reconstitute with 200µl of sterile water. Please note that if you receive this product in liquid form it has already been reconstituted as described and no further reconstitution is necessary.
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Light chain type

    unknown
  • Research areas

    • Immunology
    • Innate Immunity
    • Chemokines
    • Alpha Chemokines (CXC)
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Chemokines
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant human MCP1 protein (ab9670)
  • Recombinant Protein

    • Recombinant human MCP1 protein (ab73866)
  • sELISA pair antibody

    • Anti-MCP1 antibody (ab9858)

Applications

The Abpromise guarantee

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 (3)
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 - 2 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 ab271250 as a detection antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant MCP-1.

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 - 2 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 ab271250 as a detection antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant MCP-1.

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.
  • Target information above from: UniProt accession P13500 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6347 Human
    • Omim: 158105 Human
    • SwissProt: P13500 Human
    • Unigene: 303649 Human
    • 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

    • Western blot - Anti-MCP1 antibody (ab9669)
      Western blot - Anti-MCP1 antibody (ab9669)

      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.

    • Sandwich ELISA - Anti-MCP1 antibody (ab9669)
      Sandwich ELISA - Anti-MCP1 antibody (ab9669)
      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.
    • Western blot - Anti-MCP1 antibody (ab9669)
      Western blot - Anti-MCP1 antibody (ab9669)
      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
    • Western blot - Anti-MCP1 antibody (ab9669)
      Western blot - Anti-MCP1 antibody (ab9669)Image courtesy of Dr Laura Toma by Abreview.
      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

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (65)

    Publishing research using ab9669? Please let us know so that we can cite the reference in this datasheet.

    ab9669 has been referenced in 65 publications.

    • Sindhu S  et al. Enhanced Adipose Expression of Interferon Regulatory Factor (IRF)-5 Associates with the Signatures of Metabolic Inflammation in Diabetic Obese Patients. Cells 9:N/A (2020). PubMed: 32188105
    • Zhou DY  et al. In silico prediction and validation of potential therapeutic genes in pancreatic ß-cells associated with type 2 diabetes. Exp Ther Med 20:60 (2020). PubMed: 32952650
    • Brusletto BS  et al. Extensive Changes in Transcriptomic "Fingerprints" and Immunological Cells in the Large Organs of Patients Dying of Acute Septic Shock and Multiple Organ Failure Caused by Neisseria meningitidis. Front Cell Infect Microbiol 10:42 (2020). PubMed: 32154187
    • Lin H  et al. Silencing of long non-coding RNA Sox2ot inhibits oxidative stress and inflammation of vascular smooth muscle cells in abdominal aortic aneurysm via microRNA-145-mediated Egr1 inhibition. Aging (Albany NY) 12:12684-12702 (2020). PubMed: 32629426
    • Zhang H  et al. Long noncoding RNA MIAT2 alleviates lipopolysaccharide-induced inflammatory damage in WI-38 cells by sponging microRNA-15. J Cell Physiol 235:3690-3697 (2020). PubMed: 31566734
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 16 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MCP1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 18 2017

    Western blot abreview for Anti-MCP1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Dr. Anca Sima

    Verified customer

    Submitted Jun 18 2015

    Question

    To validate my my microarray data, I purchased MCP-1 ab (ab 9669)for IHC recently and I have been testing it on

    endometrium samples (paraffin) several times. I have tested it with several concentrations twice

    (1:100. 1:500, 1:1000) and seems like the antibody is not working for me.

    I made sure that I have right secondary, did the blocking and I tried the ab with and w/o antigen retrieval.



    Would you have any technical suggestions?

    How does your guarantee work?

    Would you be able to give me another MCP1-antibody for testing?


    Read More

    Abcam community

    Verified customer

    Asked on Aug 10 2012

    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

    Abcam Scientific Support

    Answered on Aug 10 2012

    Question

    thank you for updating me. Too bad ab73680 is out of stock. Unfortunately ab9858 is of less use to us since we can only use it for IHC-P (and ELISA) but no other techniques.
    Is it possible to get some sort of voucher instead of a replacement or refund for this antibody? We do order many antibodies at abcam so eventually we could use the voucher to buy another antibody someday. Whatever is easier for you, refund or voucher, is fine with me. Please let me know what you prefer.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 15 2012

    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

    Abcam Scientific Support

    Answered on Mar 15 2012

    Question

    And I wondered, did you already send the 'new' abcam antibodies (the replacements for MCP1 and Cx3cr1)?
    Please keep me updated on the situation; as will I. And hopefully we can take care of it soon.

    I'm sorry for all the trouble. We very much appreciate you helping us!!

    Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 14 2012

    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

    Abcam Scientific Support

    Answered on Mar 14 2012

    Question

    thank you for the update. Unfortunately I haven't heard anything from BioConnect either. I forwarded you message to my colleague, since he was the one who contacted BioConnect a few weeks ago about this.. maybe he has heard something..

    I will keep you updated and as soon as I have the information on who the contactperson was/is at BioConnect I will let you know.


    Have a nice weekend,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 02 2012

    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

    Abcam Scientific Support

    Answered on Mar 02 2012

    Question

    My apologies for the delayed reply. My colleague who has the order information contacted Bio-Connect, since the abs were indeed ordered through this distributor. Bio-Connect offered to further take of it, so they should contact you anytime now.
    Please let me know if you don't hear anything from Bio Connect, because then we will contact them again.
    Hope they have all the information you need.

    Concerning the MCP-1 antibody, we do have ab9669! I assume Bio-Connect can explain this..

    Thank you again for all the help,
    best regards

    Read More

    Abcam community

    Verified customer

    Asked on Mar 02 2012

    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

    Abcam Scientific Support

    Answered on Mar 02 2012

    Question

    My apologies for the delayed reply. My colleague who has the order information contacted Bio-Connect, since the abs were indeed ordered through this distributor. Bio-Connect offered to further take of it, so they should contact you anytime now.
    Please let me know if you don't hear anything from Bio Connect, because then we will contact them again.
    Hope they have all the information you need.

    Concerning the MCP-1 antibody, we do have ab9669! I assume Bio-Connect can explain this..

    Thank you again for all the help,
    best regards

    Read More

    Abcam community

    Verified customer

    Asked on Feb 23 2012

    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

    Abcam Scientific Support

    Answered on Feb 23 2012

    Question

    Thank you again for your quick reply and the antibody information on Cx3Cr1. We would like to replace anti-human Cx3Cr1 (ab8021) with ab8020, which you proposed.
    So, summarized:
    for anti-human TLR4 (ab22048) and anti-human TLR2 (ab9100) we would like a refund.
    And regarding the other two abs, we would like to replace them with Anti-MCP1 (ab73680) and anti-Cx3Cr1 (ab8020).

    If you need further information, please let me know!

    Best regards and enjoy the weekend,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 17 2012

    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

    Abcam Scientific Support

    Answered on Feb 17 2012

    Question

    thank you again for your quick respons! Me and my colleagues have discussed the options you gave me and decided the following.
    Concerning anti-human MCP-1 (ab9669), we would like to replace it witha different abcam antibody:
    Anti-MCP1 (ab73680)
    Although there are no references of literature on this antibody, the example image looks very specific.
    We are looking for antibodies, directed against human species and paraffin embedded material. If you have any other suggestion, feel free to let me know.

    Concerning anti-human Cx3Cr1 (ab8021), anti-human TLR4 (ab22048) and anti-human TLR2 (ab9100), we have decided to ask forthe refund. We already found replacements for TLR2 and TLR4 (from another lab). Unfortunately I could not find a proper replacement for Cx3Cr1 at abcam; Is there indeed no other anti-human Cx3Cr1 available?

    Thank you again for all the help! If you need more information, please let me know.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 16 2012

    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

    Abcam Scientific Support

    Answered on Feb 16 2012

    1-10 of 16 Abreviews or Q&A

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