Anti-MCP1 antibody [2H5] (ab21396)

Reviews (1)Q&A (8)References (10)
ELISA - Anti-MCP1 antibody [2H5] (ab21396)

    Key features and details

    • Armenian hamster monoclonal [2H5] to MCP1
    • Suitable for: ELISA
    • Reacts with: Recombinant fragment
    • Isotype: IgG

    Overview

    • Product name

      Anti-MCP1 antibody [2H5]
      See all MCP1 primary antibodies

    • Description

      Armenian hamster monoclonal [2H5] to MCP1

    • Host species

      Armenian hamster

    • Specificity

      This antibody is specific to mouse MCP-1.

    • Tested Applications & Species

      Application Species
      ELISA
      Recombinant fragment
      See all applications and species data

    • Immunogen

      CHO expressed, recombinant mouse MCP-1.

    • Positive control

    • General notes

      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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

      One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

      Learn more here.

    Properties

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab21396 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Guaranteed

    Tested applications are guaranteed to work and covered by our Abpromise guarantee.

    Predicted

    Predicted to work for this combination of applications and species but not guaranteed.

    Incompatible

    Does not work for this combination of applications and species.

    Application Species
    ELISA
    Recombinant fragment
    Application Abreviews Notes
    ELISA (1)
    Use a concentration of 5 - 10 µg/ml. To obtain a linear standard curve, serial dilutions of MCP-1 recombinant protein ranging from 2000 to 15 pg/ml are recommended for each ELISA plate.
    Notes
    ELISA
    Use a concentration of 5 - 10 µg/ml. To obtain a linear standard curve, serial dilutions of MCP-1 recombinant protein ranging from 2000 to 15 pg/ml are recommended for each ELISA plate.

    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

    • 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

    • ELISA - Anti-MCP1 antibody [2H5] (ab21396)
      ELISA - Anti-MCP1 antibody [2H5] (ab21396)
      Ab21396 staining human MCP-1 by ELISA. Image shows a standard curve obtained with increasing concentrations of human MCP-1.

    References (10)

    Publishing research using ab21396? Please let us know so that we can cite the reference in this datasheet.

    ab21396 has been referenced in 10 publications.

    • Giri J  et al. CCL2 and CXCL12 Derived from Mesenchymal Stromal Cells Cooperatively Polarize IL-10+ Tissue Macrophages to Mitigate Gut Injury. Cell Rep 30:1923-1934.e4 (2020). PubMed: 32049021
    • Yang X  et al. Metformin prevents nephrolithiasis formation by inhibiting the expression of OPN and MCP-1 in vitro and in vivo. Int J Mol Med 43:1611-1622 (2019). PubMed: 30720053
    • Lai XL  et al. Apc gene suppresses intracranial aneurysm formation and rupture through inhibiting the NF-?B signaling pathway mediated inflammatory response. Biosci Rep 39:N/A (2019). PubMed: 30808715
    • Yang SY  et al. Downregulation of angiotensin type 1 receptor and nuclear factor-?B by sirtuin 1 contributes to renoprotection in unilateral ureteral obstruction. Sci Rep 6:33705 (2016). PubMed: 27659793
    • Li X  et al. Calcium Oxalate Induces Renal Injury through Calcium-Sensing Receptor. Oxid Med Cell Longev 2016:5203801 (2016). PubMed: 27965733
    • Zhang S  et al. Comparison of various niches for endothelial progenitor cell therapy on ischemic myocardial repair: coexistence of host collateralization and Akt-mediated angiogenesis produces a superior microenvironment. Arterioscler Thromb Vasc Biol 32:910-23 (2012). PubMed: 22328781
    • Negishi T  et al. Diphenylarsinic acid increased the synthesis and release of neuroactive and vasoactive peptides in rat cerebellar astrocytes. J Neuropathol Exp Neurol 71:468-79 (2012). PubMed: 22588385
    • Zhang Y  et al. Murine sclerodermatous graft-versus-host disease, a model for human scleroderma: cutaneous cytokines, chemokines, and immune cell activation. J Immunol 168:3088-98 (2002). PubMed: 11884483
    • Luo Y  et al. Chemokine amplification in mesangial cells. J Immunol 163:3985-92 (1999). PubMed: 10491001
    • Luo Y  et al. Serologic analysis of the mouse beta chemokine JE/monocyte chemoattractant protein-1. J Immunol 153:3708-16 (1994). Inhibition Assay . PubMed: 7523503

    Customer reviews and Q&As

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

    1-9 of 9 Abreviews or Q&A

    Question

    Reference using ab21396 in IHC-Fr

    Read More
    Answer

    Below is a link to a paper that uses clone 2H5 (same clone as ab21396) for acetone fixed, frozen tissue staining (Figure 8).

    http://ajprenal.physiology.org/content/280/2/F343

    Read More

    Question

    This customer already used ab21396 (Anti-MCP1 antibody [2H5]) to conducted the wb with rat bladder tissue sample, and the results shows strong signal at 25kDa, and the references also show in the 25kDa, however, our website datasheet indicated the predicted molecular weight should be at 11kDa, therefore, she wants to know which one is the major band size? I attached her experiment image in this letter.

    Reference: Multi-analyte profiling reveals matrix metalloproteinase-9 and monocyte chemotactic protein-1 as plasma biomarkers of cardiac aging.
    http://www.ncbi.nlm.nih.gov/pubmed/21685172 2011 Aug 1;4(4):455-62. Epub 2011 Jun 17.

    Would you please help this customer to solve the problem or offer her any suggestions?

    Thanks for your kindly help

    Best regards,

    Read More
    Answer

    Thank you for contacting us.

    As shown on the following SwissProtlink, http://www.uniprot.org/uniprot/P14844, the calculated molecular weight of the Rat MCP1 is 16.4 kDa, not 11 kDa as for Human. The differences in the molecular weight observed with WB can be due to post transcriptional modifications such as glycosylation.

    I contacted the source of the antibody whoinformed me thatthey have not validated the product in house for western blot. The only data they have for the use of this clone [2H5] in Western Blot come from a publication by Luo, Y. et al., 1999, J. Immunol. 163:3985.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    1) Wo bekomme ich einen sekundaeren AK fuer Armenian Hamster her? WB, FlowCyt, ICC/IF?
    2) Gibts den auch Fluorochrom-konjugiert?

    Read More
    Answer

    Vielen Dank für Ihre Anrufe.
    Wie besprochen können wir den MCP1 Antikörper für Western blot und Durchflußzytometrie garantieren (www.abcam.com/Abpromise). In der Immuncytochemie/Immunfluoreszenz (ICC/IF) wurde er noch nicht getestet, aber es bestehen gute Chancen, dass er funktioniert, da er ja für die Immunhistochemie getestet ist.
    DISCOUNT CODE: x
    Ablaufdatum: ddmm.2012
    Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper Ab21396 in der ICC/IF zu testen, annehmen möchten. Wie am Telefon besprochen, können Sie den Gutscheincode zum Kauf eines weiteren Antikörpers einsetzen, sobald Sie uns ein Abreview für diesen Antikörper eingereicht haben. Der Discountcode hat die Nummer 100%ABR-T7ATQ und kann zum Kauf weiterer Produkte eingesetzt werden.
    Der Code wird gültig, sobald Sie uns ein Abreview für diesen Antikörper über den Test in ICC/IF eingereicht haben. Bitte geben Sie diesen Code auch im Abschnitt “Additional Notes” des Abreviews mit an, so dass wir wissen, dass sich dieses Abreview auf die Gutscheinaktion bezieht. Der Code wird dadurch aktiviert. Bitte kontaktieren Sie bei der nächsten Bestellung unsere Kundendienst mit dem Code und der ursprünglichen Bestellnummer.
    Die sekundären anti-Armenian Hamster Antikörper, die wir haben sind:
    ab5738 (unkonjugiert):
    https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-ab5738.html https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-ab5738.html.
    ab5744 (biotin):
    https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-Biotin-ab5744.html https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-Biotin-ab5744.html.
    ab5745 (HRP):
    https://www.abcam.com/Rabbit-Armenian-hamster-IgG-HL-HRP-ab5745.html https://www.abcam.com/Rabbit-Armenian-hamster-IgG-HL-HRP-ab5745.html.
    ab5746 (AP) :
    https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-Alkaline-Phosphatase-ab5746.html https://www.abcam.com/Goat-Armenian-hamster-IgG-HL-Alkaline-Phosphatase-ab5746.html.
    Falls Sie an Fluoreszenz-Konjugaten interessiert sind, sind unsere Easy Link-Kits vielleicht etwas für Sie: damit kann jeder Antikörper schnell und einfach mit einem Fluorochrom konjugiert werden. Einzelheiten finden Sie unter: www.abcam.com/Easylink
    Bitte zögern Sie nicht, sich wieder an uns zu wenden, falls Sie weitere Fragen haben. Wir freuen uns auf Ihr Abreview, egal ob Ihre Ergebnisse positiv oder negativ sind und wünschen Ihnen viel Glück für Ihre Forschung.
    Die Bedingungen dieses Angebotes befinden sich unter dem folgenden Link: www.abcam.com/collaborationdiscount
    PS: Die Bestellung (#1064638) wurde wie gewünscht storniert.

    Read More

    ELISA abreview for Anti-MCP1 antibody [2H5]

     Excellent
    Abreviews
    Application
    ELISA
    Sample
    Human Cell (aotic endothelial cells)
    Specification
    aotic endothelial cells
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Type
    Indirect
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 31 2012

    Question

    Thank you for your help.The translation: Human umbilical cord endothelial cells and smooth muscle cells in rats are right.And the customerconfirmed that MCP-1 is indeed expressed by these tissues,but they didnot get good results with the ab21396 which has seriously affected their schedule,meanwhile they thought there is no necessary to do a positive coontrol,they want a new vialof antibody to try again. So,please help me to process it,thank you very much.

    Read More
    Answer

    Thank you for kindly confirming these details as this enables us to closely monitor the quality of our products. I am glad to learn that the customer expects expression of the protein in these tissues, because following good scientific practise a positive control is paramount which is why reviewers certainly will request it. As the protocol seems absolutely fine to me, I would be pleased to arrange a replacement product free of charge with a new vial of ab21396. I am forwarding these details on to our Distributor’s Team who would be happy to organize this. Thank you for your help and cooperation. Please do not hesitate to contact us if you need anything further.

    Read More

    Question

    Dear scientist: My customer used ab21396 ( Lot. #GR37-1 ) in WB assay, but they didn't get a satisfactory result. The details listed as below. 1. Order details: · Batch No: GR37-1 · Purchase No or order No: PO#110914-1 and order no: 949109 · Antibody storage conditions (temperature/reconstitution etc) Aliquoted and stored at 4℃ 2. Please describe the problem (high background, wrong band size, more bands, no band etc).No band 3. On what material are you testing the antibody in WB? · Species: 人脐带内皮细胞和大鼠平滑肌细胞 · Cell extract or Nuclear extract: Cell extract 4. The lysate How much protein was loaded: 40ug-80ug What lysis buffer was used: RIPA What protease inhibitors were used: PMSF What loading buffer was used: 5×loading buffer Did you heat the samples: temperature and time: 100℃ 10min 5. Electrophoresis/Gel conditions/ Transfer conditions Gel percentage : 12%-15% Transfer conditions: 300MA 40-60min 6. Blocking conditions Buffer: PBST,TBST Blocking agent: milk, BSA, serum, what percentage: milk or BSA Incubation time: 2h Incubation temperature: TR 7. Primary Antibody · At what dilution(s) have you tested this antibody: 1:500 · What dilution buffer was used:PBST or TBST · Incubation time: 4℃overnight · Incubation temperature: 4℃ · What washing steps were done: PBST or TBST 10minX3or4times 8. Secondary Antibody Specification (in which species was it raised against)?.rabbit to Armenian Hamster At what dilution(s) have you tested this antibody: 1:1000;1:2000 Incubation time: 2-3h Wash steps: PBST or TBST 10minX3 or 4times Do you know whether the problems you are experiencing come from the secondary? no. 9. Detection method ECl, ECl+, other detection method: ECL, ECL+ 10. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): yes. · Is the blocking step sufficient? Yes. · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) yes · At what size are the bands migrating? Could they be degradation products f your target? 11kd 11. Optimization attempts · How many times have you tried the Western? 10times. · Do you obtain the same results every time e.g. are background bands always in the same place? Yes, no band. · What steps have you altered? We could got good result follow the same protocol.

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear your customer has had difficulty obtaining satisfactory results from this Anti-MCP1 antibody. We have received no complaints about this product therefore I am concerned it is not working for them. The protocol indeed looks absolutely fine to me. However, to fully understand the procedure I would like to ask which material has been used. The google translator gave the translation: Human umbilical cord endothelial cells and smooth muscle cells in rats. Can you confirm this please? It would be especially important to know if it is umbilical cord or spinal cord? Also, I would appreciate if the customer could confirm that MCP-1 is indeed expressed by these tissues. To my knowledge, spinal cord tissues or mixed cortical tissues (as investigated by users of our other MCP-1 antibodies, see respective Abreviews ab25124) might be an appropriate positive control, as would be smooth muscle treated with 100µg/ml glycated LDL (ab9669). If the customer cannot get hold of these controls, I might be able to send a recombinant MCP-1 protein as positive control, such as ab9780 or ab73866. Click here (or use the following: https://www.abcam.com/Recombinant-rat-MCP1-protein-ab9780.html). Click here (or use the following: https://www.abcam.com/Recombinant-human-MCP1-protein-ab73866.html). As the antibody was purchased in September, the customer is covered by our Abpromise guarantee. Therefore if the antibody does not show any results with the positive control, I would be happy to offer a refund or replacement antibody vial. Please do not hesitate to contact me should you have additional questions.

    Read More

    Question

    I want to check that if the product ab21396 can be done mmunohistology on paraffin tissues? Thank you. Regards. Looking forwards to hearing from you soon.

    Read More
    Answer

    Thank you for your enquiry. I have checked with our laboratory and it seems that this product has not been tested in paraffin embedded sections. The application that we have stated on the datasheet refers to frozen sections, and tissue/cells grown on slides. However, this does not mean that the antibody will not work in this IHC-P application, just that we have not tested it and therefore, can not guarantee that you will get good results. If you decide to go ahead and purchase this product, we encourage you to submit an Abreview via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to researchers. We reward each Abreview with Abpoints which can be used for discounts off future purchases and gifts. To find out more about our Abreview system, please see the following URL: https://www.abcam.com/abreviews I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Customer needs IHC-frozen protocol with saponin treatment. Rat retina fixation protocol and secondary.

    Read More
    Answer

    Thank you for your enquiry. The following paper has a brief description of a procedure for staining frozen sections of rat retina with MCP-1 antibody. It is not the same antibody as our hamster antibody but the procedure is standard. The important thing is to use a buffer with 0.1% saponin for the whole procedure, because it permeabilizes membranes and allows the antibody access to the MCP-1. http://www.iovs.org/cgi/content/full/42/7/1547 For a secondary antibody, if you want use a horseradish peroxidase detection, use ab6892, goat anti-hamster IgG: https://www.abcam.com/Goat-Syrian-Hamster-IgG-HL-HRP-ab6892.html Your sections will need to be blocked against non-specific binding of the anti-MCP-1 antibody to the tissue by first incubating the sections with normal goat serum. Our standard protocol for immunostaining frozen tissue is found at a link on this page: https://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=21396&mode=problems If you want to use a fluorochrome-tagged secondary antibody (eg FITC) e-mail me back and I will recommend one of our anti-hamster conjugates. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Thank you for your responses before. I am sorry for my first unclear enquiry. I am doing immunohistochemistry ,so I just want a IHC postive control, and what tissue is the positve control in my experiment. Thanks a lot again!

    Read More
    Answer

    Thank you for your reply. A MCP-1 positive control in tissue is a bit tricky as production usually tracks with disease states (for example trauma or autoimmune disease). One of the best positive controls for MCP-1 in tissue is the central nervous system of rats or mice with experimental autoimmune encephalomyelitis (see Journal of Immunology 156:3017-3023). Other labs used ischemic muscle (see 1996 or J. Surg. Res. 134:145, 2006). If you don't have access to tissues from these disease states, you might consider an adherent cell line such as the small cell lung carcinoma cell line NCI-H69 that has been treated with a drug such as doxorubicin (see Int. J. Cancer 103:380, 2003). You can grow these cells on slides as use as positive controls for IHC pretty easily. I hope that this information helps? If you have any additional questions then please get back in contact with me.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com