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    products/primary-antibodies/cd81-antibody-m38-ab79559.pdf

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Immunology Adaptive Immunity B Cells CD
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Validated using a knockout cell line

Anti-CD81 antibody [M38] (ab79559)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (73)

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Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD81 antibody [M38] (ab79559)
  • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
  • Western blot - Anti-CD81 antibody [M38] (ab79559)
  • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
  • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)

Key features and details

  • Mouse monoclonal [M38] to CD81
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

APC APC/Cy7® Biotin FITC

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Protein
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Recombinant Human CD81 protein (ab152267)
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Anti-CD63 antibody [KILL150A] - BSA and Azide free (ab271296)

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Overview

  • Product name

    Anti-CD81 antibody [M38]
    See all CD81 primary antibodies
  • Description

    Mouse monoclonal [M38] to CD81
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to Human CD81. MOLT4 cell line

  • Positive control

    • Flow Cyt: HAP1 wild type cells. MOLT4 cells. Human peripheral blood cells; IHC-P: Human liver tissue; WB: Jurkat cell lysate.
  • General notes

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    ab79559 is purified from hybridoma culture supernatant by protein A affinity chromatography. Purity is > 95% (by SDS-PAGE).
  • Clonality

    Monoclonal
  • Clone number

    M38
  • Isotype

    IgG1
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Microbiology
    • Protein
    • Human Protein
    • Receptor

Associated products

  • Alternative Versions

    • APC/Cy7® Anti-CD81 antibody [M38], prediluted (ab200565)
    • APC Anti-CD81 antibody [M38] (ab233259)
    • Biotin Anti-CD81 antibody [M38] (ab239238)
    • FITC Anti-CD81 antibody [M38] (ab239256)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human CD81 protein (ab152267)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab79559 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)
Use a concentration of 2 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use a concentration of 1 - 4 µg/ml.

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

Notes
WB
Use a concentration of 2 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use a concentration of 1 - 4 µg/ml.

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

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.
  • Target information above from: UniProt accession P60033 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 975 Human
    • Omim: 186845 Human
    • SwissProt: P60033 Human
    • Unigene: 54457 Human
    • 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

    • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
      Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-CD81 knockout cells (red line) stained with ab79559. The cells were fixed with 4% formaldehyde (10 min)  and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab79559, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) presorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.

      A mouse IgG1 isotype control antibody  (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CD81  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

      This antibody can also be used in HAP1 cells fixed with  80% methanol (5 min), , permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD81 antibody [M38] (ab79559)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD81 antibody [M38] (ab79559)

      IHC image of CD81 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab79559, 5 µg/mL, for 15 mins 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-CD81 antibody [M38] (ab79559)
      Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
      Overlay histogram showing MOLT4 cells stained with ab79559 (red line). The cells were fixed with 4% paraformaldehyde and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79559, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MOLT4 cells fixed with methanol (5 min) used under the same conditions.

      Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
    • Western blot - Anti-CD81 antibody [M38] (ab79559)
      Western blot - Anti-CD81 antibody [M38] (ab79559)
      Anti-CD81 antibody [M38] (ab79559) at 2 µg/ml + Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate

      Secondary
      IRDye800-conjugated Anti-Mouse Secondary Antibody

      Performed under non-reducing conditions.

      Predicted band size: 25 kDa
      Observed band size: 25 kDa

    • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
      Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)

      Flow cytometry analysis using human peripheral blood cells with ab79559 at 4 μg/ml.

    • Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)
      Flow Cytometry - Anti-CD81 antibody [M38] (ab79559)

      Separation of human lymphocytes (red-filled) from neutrophil granulocytes (black-dashed) in flow cytometry analysis of human peripheral whole blood stained using ab79559 at 4 μg/ml.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (73)

    Publishing research using ab79559? Please let us know so that we can cite the reference in this datasheet.

    ab79559 has been referenced in 73 publications.

    • Wu H  et al. Extracellular-vesicles delivered tumor-specific sequential nanocatalysts can be used for MRI-informed nanocatalytic Therapy of hepatocellular carcinoma. Theranostics 11:64-78 (2021). PubMed: 33391461
    • Zhou L  et al. Exosomal LncRNA LINC00659 transferred from cancer-associated fibroblasts promotes colorectal cancer cell progression via miR-342-3p/ANXA2 axis. J Transl Med 19:8 (2021). PubMed: 33407563
    • Ke Y  et al. Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90. Stem Cell Res Ther 12:21 (2021). PubMed: 33413616
    • Shan G  et al. Downregulated exosomal microRNA-148b-3p in cancer associated fibroblasts enhance chemosensitivity of bladder cancer cells by downregulating the Wnt/ß-catenin pathway and upregulating PTEN. Cell Oncol (Dordr) 44:45-59 (2021). PubMed: 33423167
    • Wu H  et al. miR-34a in extracellular vesicles from bone marrow mesenchymal stem cells reduces rheumatoid arthritis inflammation via the cyclin I/ATM/ATR/p53 axis. J Cell Mol Med 25:1896-1910 (2021). PubMed: 33465281
    View all Publications for this product

    Customer reviews and Q&As

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

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-CD81 antibody [M38]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (3T3-L1, ID8)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    15 µg
    Treatment
    Differentiated and Non-Differentiated to Adipocytes
    Specification
    3T3-L1, ID8
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 18 2022

    Immunocytochemistry/ Immunofluorescence abreview for Anti-TAPA1 antibody [M38]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (SH-SY5Y)
    Permeabilization
    Yes - 0.2% Triton-X
    Specification
    SH-SY5Y
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Nov 09 2015

    Question

    Hi,
    Thanks for your reply. The requested details are as follows:
    antibody - ab109201
    lot - GR44956-3
    order number - BRI12178.
    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 14 2012

    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

    Abcam Scientific Support

    Answered on Mar 14 2012

    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

    Abcam community

    Verified customer

    Asked on Mar 06 2012

    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

    Abcam Scientific Support

    Answered on Mar 06 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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