Overview

  • Product name
    Anti-CD81 antibody [EPR4244]
    See all CD81 primary antibodies
  • Description
    Rabbit monoclonal [EPR4244] to CD81
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human CD81 aa 150-250. The exact sequence is proprietary.
    Database link: P60033

  • Positive control
    • WB: Ramos, JAR, U87-MG, IM-9, peripheral blood mononuclear cell (PBMC), C6, PC-12, and RAW264.7 cell lysates.
  • General notes

     

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109201 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: 26 kDa.

For unpurified use at 1/500 - 1/1000.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.
  • Target

    • Function
      May play an important role in the regulation of lymphoma cell growth. Interacts with a 16-kDa Leu-13 protein to form a complex possibly involved in signal transduction. May acts a the viral receptor for HCV.
    • Tissue specificity
      Hematolymphoid, neuroectodermal and mesenchymal tumor cell lines.
    • Involvement in disease
      Defects in CD81 are the cause of immunodeficiency common variable type 6 (CVID6) [MIM:613496]; also called antibody deficiency due to CD81 defect. CVID6 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen. The defect results from a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers of circulating B cells is usually in the normal range, but can be low.
    • Sequence similarities
      Belongs to the tetraspanin (TM4SF) family.
    • Post-translational
      modifications
      Not glycosylated.
    • Cellular localization
      Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 26 kDa cell surface protein TAPA 1 antibody
      • 26 kDa cell surface protein TAPA-1 antibody
      • 26 kDa cell surface protein TAPA1 antibody
      • CD 81 antibody
      • CD81 antibody
      • CD81 antigen (target of antiproliferative antibody 1) antibody
      • CD81 antigen antibody
      • CD81 molecule antibody
      • CD81_HUMAN antibody
      • CVID6 antibody
      • S5.7 antibody
      • TAPA 1 antibody
      • TAPA1 antibody
      • Target of the antiproliferative antibody 1 antibody
      • Tetraspanin 28 antibody
      • Tetraspanin-28 antibody
      • Tetraspanin28 antibody
      • Tspan 28 antibody
      • Tspan-28 antibody
      • Tspan28 antibody
      see all

    Images

    • All lanes : Anti-CD81 antibody [EPR4244] (ab109201) at 1/700 dilution (unpurified)

      Lane 1 : Ramos cell lysate
      Lane 2 : U87-MG cell lysate
      Lane 3 : PC-12 cell lysate
      Lane 4 : Raw264.7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 26 kDa
      Observed band size: 20 kDa
      why is the actual band size different from the predicted?

    • All lanes : Anti-CD81 antibody [EPR4244] (ab109201) at 1/2000 dilution (purified)

      Lane 1 : Ramos cell lysate
      Lane 2 : U87-MG cell lysate
      Lane 3 : PC-12 cell lysate
      Lane 4 : Raw264.7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 26 kDa
      Observed band size: 20 kDa why is the actual band size different from the predicted?

    • All lanes : Anti-CD81 antibody [EPR4244] (ab109201) at 1/1000 dilution (unpurified)

      Lane 1 : Ramos cell lysate
      Lane 2 : JAR cell lysate
      Lane 3 : U87-MG cell lysate
      Lane 4 : IM-9 cell lysate
      Lane 5 : Peripheral blood mononuclear cell (PBMC) cell lysate
      Lane 6 : C6 cell lysate
      Lane 7 : PC-12 cell lysate
      Lane 8 : RAW264.7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721)

      Predicted band size: 26 kDa

    References

    This product has been referenced in:
    • Dong H  et al. Exosome-mediated transfer of lncRNA-SNHG14 promotes trastuzumab chemoresistance in breast cancer. Int J Oncol 53:1013-1026 (2018). Read more (PubMed: 30015837) »
    • Domenis R  et al. Characterization of the Proinflammatory Profile of Synovial Fluid-Derived Exosomes of Patients with Osteoarthritis. Mediators Inflamm 2017:4814987 (2017). WB ; Human . Read more (PubMed: 28634420) »
    See all 7 Publications for this product

    Customer reviews and Q&As

    1-10 of 15 Abreviews or Q&A

    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (Gradient Gel)
    Sample
    Hamster Cell lysate - whole cell (CHO cells)
    Specification
    CHO cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Mrs. Soraia Martins

    Verified customer

    Submitted Mar 16 2015

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (12%)
    Sample
    Human Cell lysate - whole cell (Cancer cell)
    Specification
    Cancer cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Dr. Hyungtaek Jeon

    Verified customer

    Submitted Nov 06 2014

    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Sample
    Human Serum (Purified exosomes from human serum)
    Specification
    Purified exosomes from human serum
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jun 20 2014

    Application
    Western blot
    Loading amount
    1e+006 cells
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Sample
    Mouse Cell lysate - whole cell (mouse embryonic stem cell)
    Specification
    mouse embryonic stem cell
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

    Abcam user community

    Verified customer

    Submitted Jan 21 2014

    Question


    The images that I send to you are T cells which express tones of CD81 that is why I didn't send any picture of HeLa cells (though based on our experience one can detect CD81 in HeLa cells).
    The problem that I am having with the rabbit Ab is that he doesn't correspond to CD81 staining in T cells. CD81 is mostly express at the pm in those cells and I have been working to understand CD81 down regulation in HIV-1 infected cells.
    IF staining protocol (standard condition):
    Briefly, CEMss cells (T cells) have been allowed to stick on poly-L-lysine treated MatTek dishes for 2h then wash once with PBS, fix using PFA 4% (wash then) for 10 min at 37 deg C, permeabilized (PBS + 1% BSA + 0.2% Triton) for 20 min at 37 deg C, primary Ab were incubated 1h at RT (rabbit anti-CD81 diluted 100 times, while mouse anti-CD81 diluted 500 times), wash , incubated with secondary Ab at RT for 30 min, wash, 2 ml PBS. Then I analyzed my cells on a deconvolution microscope.
    Western Blot (standard condition):
    When I used the Ab for WB application, I used CEMss lysates (T cells line lysed in PBS-1% Brij 96/97 containing PI) (infected or not). Using your Ab I couldn't see anything but when reprobing using our anti-CD81 I would be able to see the bands … (Conditions: 10% SDS-acrylamide gel, migration 120 V (mini-Protean system), semi-dry transfer (25V for 1h30) onto PVDF membrane, wash once TBS-T, Block in 5% milk in TBS-T, primary ON at 4 deg C, was 3 times, secondary HRP for 30 min at RT, wash 3 times, ECL.
    I also was doing IP of CD81 and I couldn't see anything either (I was using the rabbit Ab at a dilution of 1/1000). I was IPing using the ms anti-CD81 and try to detect with the Rabbit again nothing clear in my hands.
    Let me know if you have more questions/concerns based on my protocol.
    Thank you for your reply.
    Best regards,

    Read More
    Answer

    Thank you for your reply and protocol information.
    If we cannot get this antibody to work as stated on the datasheet we will replace it with a new vial of the same antibody or a different antibody of your choosing free of charge as indicated in our AbPromise guarantee. Given that, I have some more questions and protocol tips.
    Are you reducing and denaturing your sample prior to Western blot? We recommend boiling for 5 minutes at 95oC in beta-mercaptoethanol or DTT + SDS prior to loading. Since there is a disulfide bond within the residues that are recognized by this antibody, if that disulfide bond is not broken via reduction/denaturation the antibody may not properly recognize its epitope.
    For IF: In our experience testing our antibodies, Triton-X-100 is not appropriate for the use of membrane-associated antigens (such as CD81) since it destroys membranes. Since the specific epitope for ab109201 is extracellular, you should not have to permeabilize the cell membrane in order for the antibody to detect its antigen.
    For IP: As stated on the datasheet, this antibody is unsuitable for flow cytometry, IHC-P, and IP, explaining why it did not work in IP for you. The unsuitability of this antibody in these applications where CD81 is in a native conformation is most likely due to the fact that the antibody was raised against a synthetic peptide corresponding to an extracellular domain of CD81 that contains a disulfide bond (forming a loop) that is harder for the antibody to recognize in native 3D form compared to a linearized (ie. reduced, denatured for WB) form.
    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

    Answer

    Thank you for contacting Abcam.
    I am sorry to hear that this antibody is not working for you. As this was a free antibody given through our RabMab promotion, we cannot issue a refund in this case. However, if we cannot work with you to get ab109201 to work as indicated on the datasheet we will happily send you a free of charge replacement of the same antibody or another of your choosing.
    Thank you kindly for including the ppt file with your IF results. My main concern is with the lack of a band in the WB, since there is definitely a positive signal in your IF, just not colocalized with the other CD81 antibody you used. Since these two antibodies were raised against different immunogenic sequences for the same CD81 protein, it may be that some cells express CD81 in a form more recognizable by one antibody over the other.
    Thus, in order to help troubleshoot these problems, could you provide details regarding your sample, protocol and troubleshooting steps that have been taken, specifically for WB. These will not only help me to fully understand the problem but are essential should further testing of this product be needed as well. Could you also provide the Abcam order confirmation number or the PO and date of purchase for this product?
    Based on the information you have provided, I have a few recommendations. According to the Human Protein Atlas database for CD81 (http://www.proteinatlas.org/ENSG00000110651/cell), HeLa cells have very weak, if any, expression of CD81. The positive control cell lines that we have tested are Ramos, JAR, U87 MG, IM 9, Peripheral blood mononuclear cell (PBMC), C6, PC 12, and RAW264.7 cell lysates. I highly recommend trying one of these positive controls. I look forward to your reply. Please do not hesitate to contact us if you need any more advice or information.
    Below is a link with further troubleshooting steps if you have not performed them already:
    https://www.abcam.com/index.html?pageconfig=resource&rid=11352
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Question

    Catalog number(s): 109201
    Product(s): Rabbit monoclonal [EPR4244] to TAPA1

    Survey results
    ----------------------------------------------------------------------------------
    Telephone section
    Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
    A. Yes
    Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
    A. Yes

    Protocol advice section
    Q. Did Abcam's Scientific Support give you protocol advice?
    A.Yes
    Q. How did you feel about being given protocol advice?
    A.I was not expecting to be given protocol advice, but was happy to receive this
    Q. Did the product work successfully after following the advice?
    A.No
    Q. Why have you not tried the protocol advice? (Select all that apply.)
    A.

    Free of charge replacement section
    Q. Did Abcam's Scientific Support give you a free of charge replacement?
    A. Yes

    Q. Did the replacement product work successfully?
    A.Yes
    Credit note / refund section
    Q. Did Abcam's Scientific Support give you a credit note or a refund?
    A.Yes

    Q. Would you have preferred a different outcome from receiving a credit note or refund?
    A.No
    Overall satisfaction section
    Q. How satisfied are you with how the complaint was handled?
    A.Very satisfied
    Q. Could we have done anything to better resolve the problem?
    A.
    Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
    A.No, I would not mind
    Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.
    A.

    Read More
    Answer

    Thank you for your participation in Abcam's post complaint survey.

    Please let me know if there is anything else I can do for you.

    Also, if you have further questions to any of our products (e.g. primary antibodies, lysates, secondary antibodies, kits, controls, gels, proteins/peptides, slides, biochemicals, conjugation kits), please let me know as well.

    Read More

    Answer

    Vielen Dank für Ihre Antwort.

    Es tut mir leid, dass Sie Probleme mit diesen beiden Antikörpern hatten. Wie per Emailbesprochen, habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer xxx fuer ab7090 und ab93485, und sollte diese Woche bei Ihnen ankommen.

    Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Answer

    Vielen Dank fuer Ihre email.

    Ich habe nach Sekundaerantikoerpern in unserem Katalog geschaut. Wir haben zahlreiche, aber die folgenden haben wir momentan im Lager:

    ab16284 donkey anti-rabbit, HRP(https://www.abcam.com/Rabbit-IgG-secondary-antibody-ab16284.html)

    ab7090: goat anti-rabbit, HRP pre-absorbed (https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-Pre-Adsorbed-ab7090.html)

    ab6795: sheep anti-rabbit, HRP (https://www.abcam.com/Rabbit-IgG-secondary-antibody-H-L-ab6795.html)

    Alle diese Antikoerper waeren geeignet fuer den Primaerantikorper ab93485.

    Bitte sagen Sie mir doch Bescheid welchen Sekundaerantikoerper Sie bevorzugen wuerden, so dass ich die Ersatzlieferung arrangieren kann.

    Vielen Dank, und ich freue mich wieder von Ihnen zu hoeren.

    Read More

    Answer

    Vielen Dank fuer Ihre email.

    In diesem Falle koennte ich Ihnen auch einen Ersatz fuer den Primaerantikoerper anbieten.

    Wuerden Sie einen anderen C81 Antikoerper probieren wollen, da wir kein neues Batch fuer ab109201 in Lager haben? Zum Beispiel:

    ab79559 (https://www.abcam.com/TAPA1-antibody-M38-ab79559.html)
    ab93485 (https://www.abcam.com/TAPA1-antibody-ab93485.html)
    ab35026 (https://www.abcam.com/TAPA1-antibody-1D6-Azide-free-ab35026.html)
    ab128202 (https://www.abcam.com/TAPA1-antibody-ab128202.html)

    Gemaess Ihrer Wahl des Primaerantikoerpers, koennen wir dann einen neuen Sekundaerantikoerper fuer Sier heraussuchen. Sagen Sie mir doch bitte Bescheid bezueglich Ihrer Auswahl.

    Ich freue mich wieder von Ihnen zu hoeren.

    Read More

    1-10 of 15 Abreviews or Q&A

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