Recombinant
RabMAb

Recombinant Anti-CD81 antibody [EPR4244] (ab109201)

Rabbit recombinant monoclonal CD81 antibody [EPR4244]. Validated in WB and tested in Mouse, Rat, Human. Cited in 7 publication(s). Independently reviewed in 4 review(s).

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:
    • Im EJ  et al. Sulfisoxazole inhibits the secretion of small extracellular vesicles by targeting the endothelin receptor A. Nat Commun 10:1387 (2019). Read more (PubMed: 30918259) »
    • Yu J  et al. Detection of Exosomal PD-L1 RNA in Saliva of Patients With Periodontitis. Front Genet 10:202 (2019). Read more (PubMed: 30923536) »
    See all 16 Publications for this product

    Customer reviews and Q&As

    11-15 of 15 Abreviews or Q&A

    Answer

    Vielen Dank fuer Ihre email und die extra Informationen.

    Ja, Sie haben Recht - die Bande bei 26 kDa ist nicht zu sehen. Das Protein CD81 ist ein Membranprotein mit 4 helikalen Domainen. Hat der Transfer funktioniert?

    Ich wuerde vorschlagen, bis zu max. 0.1% SDS zum Transferbuffer hinzuzugeben, da das fuer hydrophobe Proteine hilfreich sein kann und es verhindert, dass diese in der Membran schon prezipitieren.

    Ein anderer Vorschlag waere (falls Sie das nicht schon gemacht haben), den Transfer mit Ponceau S (fuer die Membran) und Coomassie Blue (fuer das Gel) zu ueberpruefen. Entweder das Gel oder die Membran sollte wenigstens eine Bande bei 26 kDa zeigen.

    Ich nehme an, dassCD81 in humanen PBMCs expressiert ist?

    Die anderen Banden, die Sie detektiert haben, scheinen vom Sekundaerantikoerper zu stammen. Diesen Antikoerper kann ich Ihnen auf jeden Fall kostenlos ersetzen.

    Sagen Sie mir doch bitte Bescheid bezueglich der Fragen, und dann koennen wir entscheiden wie wir mit dem Primaerantikoerper verfahren werden.

    Ich freue mich wieder von Ihnen zu hoeren und Ihnen bei der Loesung dieses Problems zu helfen.

    Read More

    Answer

    Thank you for getting back to me and for confirming the order details and the batch number.
    I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
    Though, you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.
    Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.
    I am particularly interested in the followings:
    - sample preparation,
    - lysis buffer,
    - positive control,
    - loading control
    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

    Read More

    Answer

    Vielen Dank fürIhre Anfrage.
    Es tut mir leid, dassdiese Antikoerpern IhnenProbleme bereiten. Ich vermute, dass das Problem beim Sekundaerantikorrper liegt.

    Ich hab noch ein paar Fragen an Sie:

    1) Was fuer Proben benutzen Sie (Zell/Gewebe-type, Spezies)?
    2) Haben Sie mal probiert mit BSA zu blocken?
    3) Haben Sie auch eine noch hoehere Verduennung des Sekundaerantikoerpers versucht (z.B. 1/20,000 oder 1/50'000)?
    4) Tritt das gleiche Problem auch mit anderen Primaerantikoerpern auf?
    5) Was ist Ihre Bestellnummer?

    Die Bande um die 50-60 kDakoennte endogenes IgG (heavy chain) sein - falls Sie mit Gewebeproben arbeiten.

    Der Sekundaerantikoerper sollte eigentlich kein Signal geben wenn der Primaerantikoerper weggelassen wurde.

    Ich freue mich wieder von Ihnen zu hoeren und Ihnen bei der Loesung dieses Problems zu helfen.

    Read More

    Question

    Hi there,



    A couple of months ago I purchased your rabbit anti-CD81 (anti-TAPA-1) antibody (ab109201) which is guaranteed to detect human CD81 by western blot. I have been unable to detect CD81 in blots and wonder if I am doing something incorrectly as another one of your antibodies (ab79559) also failed to wprk in western blot.



    I am using 2 cell lines (Huh7.5 and 293T cells) and I have shown by flow cytometry that both contain surface CD81:

    - they both strongly bind another one of your anti-CD81 antibodies (ab79559 - flow cytometry and WB guaranteed);

    - they both bind HCV E2 (the receptor of which is CD81);

    - cells pre-incubated withab79559 no longer bind HCV E2 (as CD81 is blocked by your antibody).



    I wanted to confirm the presence of CD81 by WB so performed SDS-PAGE withwhole cell lysates (from Huh7.5, 293T, Raji, Jurkat and other cell lines) using standard protocols, transferred proteins onto nitrocellulose, blocked for 2 h with 10 % milk, and probed overnight withab79559 at 1:1000 dilution in 5 % milk in TBS-T, followed by anti-mouse-HRP. I even got no signal, even when reprobed at 1:100 dilution (incidentally, the problem is not with the secondary as that works with other antibodies).



    I assumed thatab79559 must detect a 'correctly folded' epitope as it works with flow cytometry and that this must be altered by SDS-PAGE so I tried your WB-specific anti-CD81 antibody (ab109201). WB was performed as above, but using an anti-rabbit-HRP secondary, and again I got no signal.





    I then transiently transfected 293T, 293T/17 and Huh7.5 cells with myc-DDK-tagged CD81 on the CMV promoter (Origene RC217508), and after 48 h expression performed SDS-PAGE and WB using whole cell lysates of transfected and non-transfected cells. I then probed one blot with your WB-guaranteed anti-CD81 antibody (ab109201) and another blot with an anti-flag antibody. The anti-flag antibody strongly detected CD81 in lanes from CD81-transfected cells, but nothing in non-transfected lanes. Strangely, your anti-CD81 antibody detected nothing - even the transfected lanes had no signal.



    I had begun to think that CD81 perhaps did not run into the gel during SDS-PAGE (as membrane proteins can sometimes be temperamental), but I am perplexed that I can detect tagged CD81 with the anti-tag antibody, but not with your anti-CD81. Please can you tell me if there is something that I have failed to do correctly for the detection of CD81 by WB so that I can test the antibody again.



    Kind regards,

    Read More
    Answer

    Thank you for your enquiry regarding ab109201 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.
    As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
    Could you please provide the batch and order numbers for ab109201?
    I look forward to hearing from you and hope to solve this problem as soon as possible.

    Read More

    Answer

    Thank you for your message. I am sorry it has taken some time to confirm this information for you. Reviewing all the TAPA-1 antibodies we have on the catalog, most of them are raised against a cell line, which means the antibody could be detecting any region of the full length protein. I am sorry that these are regrettably not eptitope mapped.  We do have the following two antibodies which have more specific immunogen sequences, as they have been raised against peptide. I can suggest to review the online datasheets for further information and to ensure to select one that is tested and therefore guaranteed in the application and species you wish to use. ab93485 Immunogen:  NKDQIAKDVKQFYDQA Amino acids 114 - 129. This is in the extracellular domain, but just out of the region you want to look at. Tested in: WB Reacts with: Hu Click here (or use the following: https://www.abcam.com/index.html?datasheet=93485). ab109201  Immunogen: Between amino acids 175 and 200 of human CD81/TAPA1 (UniProt: P60033). Tested in: WB, ICC Reacts with: Ms, Rat, Hu Click here (or use the following: https://www.abcam.com/index.html?datasheet=109201). I hope this will be helpful to you. If you require any further information, please do not hesitate to contact us.  

    Read More

    11-15 of 15 Abreviews or Q&A

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