Validated using a knockout cell line

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

Reviews (3)Q&A (5)References (22)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [MEM-M6/1] (ab666)
  • Western blot - Anti-CD147 antibody [MEM-M6/1] (ab666)
  • Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)
  • Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)
  • Flow Cytometry - Anti-CD147 antibody [MEM-M6/1] (ab666)

Key features and details

  • Mouse monoclonal [MEM-M6/1] to CD147
  • Suitable for: IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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, 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: A549 cells and Peripheral blood lymphocytes. WB: A549, Raji and Jurkat.

  • 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.

    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

Applications

The Abpromise guarantee

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 (2)
Use a concentration of 10 µg/ml.
Flow Cyt
Use a concentration of 10 µg/ml.

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

 
WB (1)
Use a concentration of 1 µg/ml. Use under non reducing condition.
Notes
IHC-P
Use a concentration of 10 µg/ml.
Flow Cyt
Use a concentration of 10 µg/ml.

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

 
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.

    • Western blot - Anti-CD147 antibody [MEM-M6/1] (ab666)
      Western blot - Anti-CD147 antibody [MEM-M6/1] (ab666)
      All lanes : Anti-CD147 antibody [MEM-M6/1] (ab666) at 1 µg/ml

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : BSG knockout A549 cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Observed band size: 55-70 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab666 observed at 55-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

      ab666 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab666 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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

      Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with ab666 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab666) (1x106 in 100μl at 10 μg/ml) for 30 min at 4°C.

      The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.

      Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

    • 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 (22)

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

    ab666 has been referenced in 22 publications.

    • Wagatsuma T  et al. Discovery of Pancreatic Ductal Adenocarcinoma-Related Aberrant Glycosylations: A Multilateral Approach of Lectin Microarray-Based Tissue Glycomic Profiling With Public Transcriptomic Datasets. Front Oncol 10:338 (2020). PubMed: 32232009
    • Colangelo NW & Azzam EI Extracellular vesicles originating from glioblastoma cells increase metalloproteinase release by astrocytes: the role of CD147 (EMMPRIN) and ionizing radiation. Cell Commun Signal 18:21 (2020). PubMed: 32033611
    • Aguiar JA  et al. Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue. Eur Respir J 56:N/A (2020). PubMed: 32675206
    • Essahib W  et al. SARS-CoV-2 host receptors ACE2 and CD147 (BSG) are present on human oocytes and blastocysts. J Assist Reprod Genet 37:2657-2660 (2020). PubMed: 32959144
    • Dhanda AS  et al. Distribution of CD147 During Enteropathogenic Escherichia coli and Salmonella enterica Serovar Typhimurium Infections. Anat Rec (Hoboken) 302:2224-2232 (2019). PubMed: 31443124
    View all Publications for this product

    Customer reviews and Q&As

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    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-CD147 antibody [MEM-M6/1]

     Good
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HT29 cells)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    20 µg
    Specification
    HT29 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 14 2021

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

     Poor
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mouse kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: commercial - ZYTOMED's OmniPrep (pH 9)
    Permeabilization
    Yes - 0.05% tween 20 in the washing buffer
    Specification
    Mouse kidney
    Blocking step
    commercial - Invitrogen's CAS-Block as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 13 2020

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

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Adult human Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: commercial - ZYTOMED's OmniPrep (pH 9)
    Permeabilization
    Yes - 0.05% tween 20 in the washing buffer
    Specification
    Adult human Kidney
    Blocking step
    commercial - Invitrogen's CAS-Block as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Yehudit Gnatek

    Verified customer

    Submitted May 11 2020

    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.

    Read More

    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.

    Read More

    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.

    Read More

    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.        

    Read More
    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. 

    Read More

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