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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD147 antibody [EPR4053] (ab108308)

Submit a review Q&A (6)References (7)
Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR4053] (ab108308)
  • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
  • Flow Cytometry - Anti-CD147 antibody [EPR4053] (ab108308)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [EPR4053] (ab108308)
  • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
  • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
  • Anti-CD147 antibody [EPR4053] (ab108308)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4053] to CD147
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

Overview

  • Product name

    Anti-CD147 antibody [EPR4053]
    See all CD147 primary antibodies

  • Description

    Rabbit monoclonal [EPR4053] to CD147

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, Raji, HEK-293T, Jurkat, HuT-78, A431, U-87 MG and HeLa cell lysates; IHC-P: Human colon cancer tissue; ICC/IF: HeLa cells; Flow Cyt: HeLa cells. WB: HeLa and U-87 MG whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108308 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
1/1000 - 1/10000. Predicted molecular weight: 42 kDa.
IHC-P
1/3000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.
Flow Cyt
1/30.
ICC/IF
1/50.

For unpurified use at 1/500.

 
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 42 kDa.
IHC-P
1/3000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.
Flow Cyt
1/30.
ICC/IF
1/50.

For unpurified use at 1/500.

 

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

    • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      All lanes : Anti-CD147 antibody [EPR4053] (ab108308) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 42 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR4053] (ab108308)
      Immunocytochemistry/ Immunofluorescence - Anti-CD147 antibody [EPR4053] (ab108308)
      Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling CD147 with purified ab108308 at 1:50 dilution (6.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor ® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor ® 488 ,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      All lanes : Anti-CD147 antibody [EPR4053] (ab108308) at 1/1000 dilution

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

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 42 kDa
      Observed band size: 42-70 kDa why is the actual band size different from the predicted?



      Lanes 1 - 3: Merged signal (red and green). Green - ab108308 observed at 42-70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab108308 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 ab108308 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-CD147 antibody [EPR4053] (ab108308)
      Flow Cytometry - Anti-CD147 antibody [EPR4053] (ab108308)
      Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CD147 with purified ab108308 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [EPR4053] (ab108308)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD147 antibody [EPR4053] (ab108308)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling CD147 with purified ab108308 at 1/3000 dilution (0.1 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      All lanes : Anti-CD147 antibody [EPR4053] (ab108308) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : BSG knockout HEK293T cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : U-87 MG cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 42 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab108308 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.


      ab108308 Anti-CD147 antibody [EPR4053] was shown to specifically react with CD147 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266331 (knockout cell lysate ab256853) was used. Wild-type and CD147 knockout samples were subjected to SDS-PAGE. ab108308 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      Western blot - Anti-CD147 antibody [EPR4053] (ab108308)
      All lanes : Anti-CD147 antibody [EPR4053] (ab108308) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : BSG (Basigin) knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : U-87 MG whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 42 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab108308 observed at 42 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab108308 was shown to specifically react with Basigin in wild-type HAP1 cells as signal was lost in BSG (Basigin) knockout cells. Wild-type and BSG (Basigin) knockout samples were subjected to SDS-PAGE. Ab108308 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Anti-CD147 antibody [EPR4053] (ab108308)
      Anti-CD147 antibody [EPR4053] (ab108308)

    References (7)

    Publishing research using ab108308? Please let us know so that we can cite the reference in this datasheet.

    ab108308 has been referenced in 7 publications.

    • Mahmoud AM & Ali MM High Glucose and Advanced Glycation End Products Induce CD147-Mediated MMP Activity in Human Adipocytes. Cells 10:N/A (2021). PubMed: 34440867
    • Song Y  et al. Inhibition of EMMPRIN by microRNA-124 suppresses the growth, invasion and tumorigenicity of gliomas. Exp Ther Med 22:930 (2021). PubMed: 34306199
    • Bu X  et al. CD147 confers temozolomide resistance of glioma cells via the regulation of β-TrCP/Nrf2 pathway. Int J Biol Sci 17:3013-3023 (2021). PubMed: 34421346
    • Ali MM  et al. CD147 Levels in Blood and Adipose Tissues Correlate with Vascular Dysfunction in Obese Diabetic Adults. J Cardiovasc Dev Dis 9:N/A (2021). PubMed: 35050217
    • Flatt JF  et al. Expression of South East Asian Ovalocytic Band 3 Disrupts Erythroblast Cytokinesis and Reticulocyte Maturation. Front Physiol 11:357 (2020). PubMed: 32411010
    • Himbert D  et al. Characterization of CD147, CA9, and CD70 as Tumor-Specific Markers on Extracellular Vesicles in Clear Cell Renal Cell Carcinoma. Diagnostics (Basel) 10:N/A (2020). PubMed: 33276608
    • Sugyo A  et al. Evaluation of (89)Zr-labeled human anti-CD147 monoclonal antibody as a positron emission tomography probe in a mouse model of pancreatic cancer. PLoS One 8:e61230 (2013). WB ; Human . PubMed: 23577210

    Customer reviews and Q&As

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

    1-6 of 6 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.

    Read More

    Question

    please accept my apologies for the late response, I attended a
    conference and was busy in the lab.
    I also tried to use both antibodies under non-reducing conditions,
    however, this did not improve the detectability of CD147 by ab108308
    (see attachment). I increased the concentration (1:500), which is now
    twice as much as the highest concentration recommended.
    Therefore, it would be very kind if you could send us a replacement. I
    attached the goods receipt slip including the order date as well as
    the order number Unfortunately,
    the antibodies were ordered on the , which,
    considering my first e-mail, is about two weeks over the 120 day
    deadline. Would it be possible to receive a replacement nevertheless?
    We bought this antibody specifically for Western blot purposes as it
    detects an intracellular epitope.
    We would me much obliged!
    Additionally, in contrast to the ab108308 data sheet, there is no
    information on the ab666 data sheet regarding cross-reactivity with
    other species. Do you have any information on that topic?

    Read More
    Answer

    Thank you for your reply.

    I would be more than happy to send you a replacement antibody. If you could let me know the catalogue number of the antibody you would like as a replacement, I will be happy to arrange that for you.

    As for ab666, we have only tested it on human samples and so not have any other data on how it would react with other species.

    Looking forward to your reply.

    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


    Please find completed from. We would like a credit note in order to buy a
    different antibody.

    Thanks,

    Order Details
    Antibody code: AB108308
    Lot number:
    - GR46693-3

    General Information
    Antibody storage conditions (temperature/reconstitution etc)
    - Aliquoted for storage at -20C with working aliquot stored at 4C
    Description of the problem (high background, low signal, non-specific
    satining etc.)
    - No signal
    Sample (Species/Tissue/Cell Type/Cell Line etc.)
    - Fresh human mature erythrocytes
    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration
    etc.)
    - 1% formaldehyde + 0.0075% glutaraldehyde
    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
    - None
    Permeabilization step
    - 0.1% Triton-X100 for 5 min
    Blocking conditions (Buffer/time period, Blocking agent etc.)
    - 0.1% BSA in PBS in all washes before (and after) antibody application
    (approx 1 hr) then antibody applied in 4% BSA (in PBS)
    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
    Wash step)
    - Abcam ab108308 anti-CD147 rabbit monoclonal antibody 1:100 for 1 hr in 4%
    BSA in PBS
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,
    Wash step)
    - Invitrogen Alexa-546 goat anti-rabbit antibody 1:250 for 30 min in 4% goat
    serum
    Detection method
    - laser scanning confocal microscopy
    Positive and negative controls used (please specify)
    - negative control; no primary antibody
    - positive controls; other in-house rabbit antibodies (eg anti-GPA) and we
    have since found anti-CD147 mouse monoclonal antibodies that work using this
    method (eg clone M6/6)
    Optimization attempts (problem solving) How many times have you tried the
    IHC?
    - 2
    Have you run a "No Primary" control?
    - Yes
    Do you obtain the same results every time?
    - Yes
    What steps have you altered?
    - None
    Additional Notes

    Read More
    Answer

    Thank you for your reply and for kindly confirming these details.

    I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for you.

    Credit ID: ####

    The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    In addition to this, having reviewed the details provided, I recommend you may like to consider the following for future experiments, which I hope would be helpful:

    1. How long was the fixation? We would recommend 10 minutes only to ensure there is not too much crosslinking.

    2. As CD147 is a cell membrane protein, permeabilization is not required. Using a strong detergent like Triton can disrupt membrane proteins and significantly affect some epitopes which can affect the results of the staining. Epitopes for individual antibodies could be affected differently, which may be why the other antibody is working.

    3. A more gentle detergent such as 0.1% Tween 20 can be used in the wash buffer and antibody dilution buffer to help keep the antibody solubilized.

    4. Overnight incubation at 4oC with primary antibody can often provide more effective and specific staining.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Question

    Dear Abcam,
    We recently purchase this antibody (PO 61343642 06/01/12). We wanted to use it to label fixed, permeabilisedred cells, visualised using confocal microscopy. The antibody was shown on your website to work for IHC, ICC and we therefore thought it would work for confocal. Unfortunately the antibody did not work.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CD147 antibody [EPR4053] (ab108308)https://www.abcam.com/CD147-antibody-EPR4053-ab108308.html#CD147-Primary-antibodies-ab108308-4.jpg

    We have a lot of experience with this technique so I don't think the failure is due to our error. We have now found an alternative antibody that does work well. We buy a lot of antibodies from your company and would (if possible) like a replacement antibody.

    Best wishes,

    Read More
    Answer

    Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful.

    I would like to reassure you that this antibody is tested and covered by our guarantee for IHC-P, ICC and human samples. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality montoring.

    In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.

    In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

    Thank you for your time and cooperation. We look forward to receiving the completed questionaire and details of how you would like to proceed.

    Order Details

    Antibody code:

    Lot number:
    Purchase order number
    or preferably Abcam order number:

    General Information

    Antibody storage conditions (temperature/reconstitution etc)
    Description of the problem (high background, low signal, non-specific satining etc.)

    Sample (Species/Tissue/Cell Type/Cell Line etc.)

    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)

    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)

    Permeabilization step

    Blocking conditions (Buffer/time period, Blocking agent etc.)

    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)

    Detection method

    Positive and negative controls used (please specify)

    Optimization attempts (problem solving)

    How many times have you tried the IHC?

    Have you run a "No Primary" control?
    Yes No

    Do you obtain the same results every time?
    Yes No

    What steps have you altered?

    Additional Notes

    We would appreciate if you are also able to provide and image which would help us to assess the results

    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