Anti-CD147 antibody [MEM-M6/1] (ab666)

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Overview

  • Product name
    Anti-CD147 antibody [MEM-M6/1]
    See all CD147 primary antibodies
  • Description
    Mouse monoclonal [MEM-M6/1] to CD147
  • Host species
    Mouse
  • Specificity
    Human CD147 antigen. This antibody recognizes an epitope in the N-terminal Ig domain (D1). This high-affinity antibody is capable of binding to unstimulated peripheral blood T cells.
  • Tested applications
    Suitable for: IHC-P, Flow Cyt, IP, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human CD147. Purified soluble recombinant form of CD147, CD147Rg, which consists of the cDNA coding for the hinge region, CH2-and CH3 domain of human IgG1 (CD147Rg is secreted by transfectants as a dimer).
    Database link: P35613

  • Positive control
    • This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle. Flow Cytometry: Peripheral blood lymphocytes.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 24th January 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab666 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
IHC-P Use a concentration of 10 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Use under non reducing condition.

Target

  • Function
    Plays pivotal roles in spermatogenesis, embryo implantation, neural network formation and tumor progression. Stimulates adjacent fibroblasts to produce matrix metalloproteinases (MMPS). May target monocarboxylate transporters SLC16A1, SLC16A3 and SLC16A8 to plasma membranes of retinal pigment epithelium and neural retina. Seems to be a receptor for oligomannosidic glycans. In vitro, promotes outgrowth of astrocytic processes.
  • Tissue specificity
    Present only in vascular endothelium in non-neoplastic regions of the brain, whereas it is present in tumor cells but not in proliferating blood vessels in malignant gliomas.
  • Sequence similarities
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications
    N-glycosylated.
  • Cellular localization
    Cell membrane. Melanosome. Colocalizes with SLC16A1 and SLC16A8 (By similarity). Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
    • Alternative names
      • 5A11 antigen antibody
      • 5F7 antibody
      • BASI_HUMAN antibody
      • Basigin (Ok blood group) antibody
      • Basigin antibody
      • Blood brain barrier HT7 antigen antibody
      • Bsg antibody
      • CD 147 antibody
      • CD147 antibody
      • CD147 antigen antibody
      • Collagenase stimulatory factor antibody
      • EMMPRIN antibody
      • Extracellular matrix metalloproteinase inducer antibody
      • Leukocyte activation antigen M6 antibody
      • M 6 antibody
      • M6 antibody
      • M6 leukocyte activation antigen antibody
      • Neurothelin antibody
      • OK antibody
      • OK blood group antibody
      • OK blood group antigen antibody
      • TCSF antibody
      • Tumor cell derived collagenase stimulatory factor antibody
      • Tumor cell-derived collagenase stimulatory factor antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [MEM-M6/1] (ab666)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [MEM-M6/1] (ab666)

      IHC image of CD147 staining in human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab666, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)
      Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)
      Flow cytometry of human peripheral blood cells with ab666 at 1 µg/ml
    • Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)
      Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)

      Overlay histogram showing peripheral blood lymphocytes stained with ab666 (red line). The cells were incubated with the antibody (ab666, 1 µg/1x106 cells) for 30 minutes at 4ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 4ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed gating on peripheral blood lymphocytes.

    References

    This product has been referenced in:
    • Badillo-Soto MA  et al. Potential protein targets of the peptidylarginine deiminase 2 and peptidylarginine deiminase 4 enzymes in rheumatoid synovial tissue and its possible meaning. Eur J Rheumatol 3:44-49 (2016). Read more (PubMed: 27708970) »
    • Huang XQ  et al. Co-expression of CD147 and GLUT-1 indicates radiation resistance and poor prognosis in cervical squamous cell carcinoma. Int J Clin Exp Pathol 7:1651-66 (2014). Read more (PubMed: 24817962) »
    See all 10 Publications for this product

    Publishing research using ab666? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Question

    many thanks for your generous offer! The antibody we require should
    have the following properties (as we would like to detect human CD147
    from xenografts grown in mice later on):
    - human-specific (must not detect mouse CD147)
    - must not be from mouse (or directly HRP-conjugated from mouse)
    - detect all 4 isoforms (bind C-terminally or on the extracellular
    domain closest to the cell membrane, which is also expressed in all
    isoforms)
    Unfortunately, as far as I can see, ab108308 is the only antibody on
    your list which fulfils all these criteria. What would you recommend?
    Try the same antibody again from another batch (maybe other users
    reported problems or malfunction with this antibody, also?) or go for
    another antibody?
    I hope we can find a satisfying solution; ab666 yielded very nice and
    promising results, however, we need an antibody which fulfils the
    criteria stated above urgently to confirm and extend our experiments.
    I am looking forward to your response!
    Again, thank you very much for your offer and have a pleasant evening!

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. It does look like ab108308 may be the best fit for your experiment, so as requested, I have issued a free of charge replacement of ab108308.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

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    Question

    please find the following protocol, if you need any more specific information, I am glad to provide it! Dilute protein (12.5 µg/slot) with RIPA buffer Add protein loading buffer (including 2% SDS and 5% beta-mercaptoethanol) Denaturation (and reduction) 5 min at 95 °C --> on ice PAGE at 70-80 V for 3h on 12% gels (I increased the voltage when it took too long) Blotting on PVDF (activation: 5 sec MeOH; 5 min H2O; 15 min blotting buffer) or nitrocellulose (activation: 15 min in 20% MeOH buffer) membrane at 350 mA for 100 min. Ponceau S & wash off in PBS Blocking in PBS/0.1% Tween 20/5% SMP for 1 h Primary antibodies: ab666 & ab108308 1:1000 (1 µg/ml each); shake at 4°C overnight Wash membranes 2x with PBS/0.1% Tween 20 Secondary antibody (HRP-conjugated) 1:3000 (protocol: 1:5000); shake 45 min at RT Wash 2x with PBS/0.1% Tween 20 & 2x with PBS Incubate 1 min with Luminol reagent; development: 10 sec or 60 sec (I sent you an image of the latter) That is it from my side, again, if I forgot to mention something important, I am happy to provide the information! Hopefully, I do not need to exchange the antibody, as it may be helpful to find splice variant basigin-3, and as a band can be observed in that region, I am very hopeful to get the protocol settled! Many thanks in advance and have a nice evening!

    Read More
    Answer

    Thank you for your message and for providing this further information.

    I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would like to offer a free of charge replacement in compensation (providing the product has been purchased in the last 120 days). In order to arrange this, I would appreciate if you could confirm your order number and date of purchase?

    Thank you for your continued cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    many thanks for your response! I performed a Western Blot to detect CD147 utilizing both antibodies ab666 and ab108308. Although I used the conditions recommended in the data sheet for ab108308 (reducing (and denaturating) conditions; 1:1000 dilution; 12.5 µg protein per lane), I could not detect anything in the range expected (and confirmed by ab666). Please find a copy of the blot attached. To the left of the marker protein ab666 was used, right of the marker ab108308. As you confirmed, ab666 detects an epitope on the second extracellular loop and therefore only splice variant basigin-2 (and basigin-1, which is retina-specific), and ab108308 detects a C-terminal epitope (and therefore all 4 known splice variants). However, as you can see, I achieved some decent results for ab666 detecting both the highly- (~50-60 kDa) and low-glycosylated (~33 kDa) forms of CD147, but nothing in that range for ab108308. The only very slight bands I was able to detect might (from the protein mass) be attributed to basigin-3 (~28 kDa). Therefore, I turn to you with the question what I might improve to improve my results and detect the same bands I got with ab666.

    Read More
    Answer

    Thank you for contacting us.


    I am sorry to hear that of this problem. No or low signal can be the result of a number of factors. As this product is a tissue culture supernatant I would suggest that prehaps increasing the antibody concentration may help. If you could send me the protocol that you have used I may be able to offer furhter tips as well.



    This product is covered by our Abpromise guarantee.If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, if being used according to specifications listed on our datasheet.



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

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    Question

    we purchased anti-human CD147 antibody clone MEM-M6/1 (ab666). However, due to different nomenclatures, I am not sure what epitope, and what isoforms / splice variants in particular, this antibody detects. With respect to the attached paper (Liao et al. 2011; Figure 1E), could you please specify which isoforms should be recognized by this antibody? Another paper (Yu et al. 2008), in which the crystal structure of basigin-2 was resolved, might also be of aid and emphasize difficulties regarding the nomenclature of domains. Many thanks in advance and a pleasant weekend!

    Read More
    Answer

    Thank you for contacting us.

    The epitope should be located within the N-teminal Ig domain D1, which is the more membrane-distal domain, thus IgC2 domain as called within the paper mentioned.The most information about epitope of thios product is contained in the following paper:Koch C, Staffler G, Huttinger R, Hilgert I, Prager E, Cerny J, Steinlein P, Majdic O, Horejsi V, Stockinger H: T cell activation-associated epitopes of CD147 in regulation of the T cell response, and their definition by antibody affinity and antigen density. Int Immunol. 1999 May;11(5):777-86.

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

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    Question

    We are interested in buying from Abcam the products [MEM-M6/1] (ab666) and [MEM-M6/6] - Low Endotoxin (ab119114).   In our experiments it is very important to know  the isotype of the light chain. May I please ask you if you know the above information.        

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    Answer

    Thank you for contacting Abcam regarding these antibodies. I have spoken with our laboratory regarding the light chain isotypes of ab666 and ab119114. Unfortunately this information is not available. Please do not hesitate to contact us if there is anything else that we may do to help you reach your research goals. 

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

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