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  1. Link

    products/primary-antibodies/cd63-antibody-nk1c3-ab1318.pdf

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Cardiovascular Blood Platelets
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Validated using a knockout cell line

Anti-CD63 antibody [NK1/C3] (ab1318)

  • Datasheet
Reviews (1)Q&A (3)References (15)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [NK1/C3] (ab1318)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
  • Flow Cytometry - Anti-CD63 antibody [NK1/C3] (ab1318)

Key features and details

  • Mouse monoclonal [NK1/C3] to CD63
  • Suitable for: ICC/IF, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-CD63 antibody [NK1/C3]
    See all CD63 primary antibodies
  • Description

    Mouse monoclonal [NK1/C3] to CD63
  • Host species

    Mouse
  • Specificity

    This antibody is specific to a 53 kD protein, expressed on activated platelets. CD63 is a lysosomal membrane glycoprotein that is translocated to plasma membrane after platelet activation. It is also expressed on granulocytes, B cells and T cells.
  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to CD63. Smooth plasma membrane fraction of MeWo cells.

  • Positive control

    • ICC/IF: HAP1 cells (HAP1-CD63 knockout cells used as negative cell line)
  • General notes


    CD63 is expressed on activated platelets. CD63 is a lysosomal membrane glycoprotein that is translocated to plasma membrane after platelet activation. It is also present in monocytes and macrophages and is weakly expressed in granulocytes, B, and T cells. CD63 is identical to the melanoma-associated antigen ME491 and to the platelet antigen PTLGP40.

    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. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.3
    Preservative: 0.05% Sodium azide
    Constituent: 1% BSA
  • Concentration information loading...
  • Purity

    IgG fraction
  • Primary antibody notes

    CD63 is expressed on activated platelets. CD63 is a lysosomal membrane glycoprotein that is translocated to plasma membrane after platelet activation. It is also present in monocytes and macrophages and is weakly expressed in granulocytes, B, and T cells. CD63 is identical to the melanoma-associated antigen ME491 and to the platelet antigen PTLGP40.
  • Clonality

    Monoclonal
  • Clone number

    NK1/C3
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Cardiovascular
    • Blood
    • Platelets
    • Immunology
    • Cell Type Markers
    • CD
    • Platelets
    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Other
    • Cancer
    • Tumor biomarkers
    • Tumor antigens

Associated products

  • Compatible Secondaries

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

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab1318 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
ICC/IF
Use a concentration of 1 - 5 µg/ml.

Signal can be observed in cells fixed with methanol or PFA.

IHC-P
1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
1/100.

(Also see PMID 19129916).

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

Notes
ICC/IF
Use a concentration of 1 - 5 µg/ml.

Signal can be observed in cells fixed with methanol or PFA.

IHC-P
1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt
1/100.

(Also see PMID 19129916).

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

Target

  • Function

    This antigen is associated with early stages of melanoma tumor progression. May play a role in growth regulation.
  • Tissue specificity

    Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.
  • Sequence similarities

    Belongs to the tetraspanin (TM4SF) family.
  • Cellular localization

    Cell membrane. Lysosome membrane. Late endosome membrane. Also found in Weibel-Palade bodies of endothelial cells. Located in platelet dense granules.
  • Target information above from: UniProt accession P08962 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 967 Human
    • Omim: 155740 Human
    • SwissProt: P08962 Human
    • Unigene: 445570 Human
    • Alternative names

      • Lysosomal associated membrane protein 3 antibody
      • CD 63 antibody
      • CD63 antibody
      • CD63 antigen (melanoma 1 antigen) antibody
      • CD63 antigen antibody
      • CD63 antigen melanoma 1 antigen antibody
      • CD63 molecule antibody
      • CD63_HUMAN antibody
      • gp55 antibody
      • Granulophysin antibody
      • LAMP 3 antibody
      • LAMP-3 antibody
      • LAMP3 antibody
      • LIMP antibody
      • Lysosomal-associated membrane protein 3 antibody
      • Lysosome associated membrane glycoprotein 3 antibody
      • Lysosome-associated membrane glycoprotein 3 antibody
      • Mast cell antigen AD1 antibody
      • ME491 antibody
      • Melanoma 1 antigen antibody
      • Melanoma associated antigen ME491 antibody
      • Melanoma associated antigen MLA1 antibody
      • Melanoma-associated antigen ME491 antibody
      • MGC72893 antibody
      • MLA 1 antibody
      • MLA1 antibody
      • NGA antibody
      • Ocular melanoma associated antigen antibody
      • Ocular melanoma-associated antigen antibody
      • OMA81H antibody
      • PTLGP40 antibody
      • Tetraspanin 30 antibody
      • Tetraspanin-30 antibody
      • Tspan 30 antibody
      • Tspan-30 antibody
      • TSPAN30 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
      Formalin-fixed, paraffin-embeded human melanoma tissue stained for CD63 using ab1318 at 1/100 dilution in immunohistochemical analysis. Antigen retrieval with citrate buffer pH 6.0
    • Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [NK1/C3] (ab1318)
      Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [NK1/C3] (ab1318)

      ab1318 staining CD63 in wild-type HAP1 cells (top panel) and CD63 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab1318 at 1ug/ml and ab6046 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in pseudo color red).  Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [NK1/C3] (ab1318)
      ab1318 staining human melanoma by IHC-P.
    • Flow Cytometry - Anti-CD63 antibody [NK1/C3] (ab1318)
      Flow Cytometry - Anti-CD63 antibody [NK1/C3] (ab1318)
      Overlay histogram showing HL60 cells stained with ab1318 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab1318, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-Mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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. This antibody gave a positive signal in HL60 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (15)

    Publishing research using ab1318? Please let us know so that we can cite the reference in this datasheet.

    ab1318 has been referenced in 15 publications.

    • Ivanusic D  et al. The large extracellular loop of CD63 interacts with gp41 of HIV-1 and is essential for establishing the virological synapse. Sci Rep 11:10011 (2021). PubMed: 33976357
    • Oh JG  et al. Analysis of extracellular vesicle miRNA profiles in heart failure. J Cell Mol Med 24:7214-7227 (2020). PubMed: 32485073
    • Albacete-Albacete L  et al. ECM deposition is driven by caveolin-1-dependent regulation of exosomal biogenesis and cargo sorting. J Cell Biol 219:N/A (2020). PubMed: 33053168
    • Yokoi A  et al. Mechanisms of nuclear content loading to exosomes. Sci Adv 5:eaax8849 (2019). PubMed: 31799396
    • Aaberg-Jessen C  et al. Co-expression of TIMP-1 and its cell surface binding partner CD63 in glioblastomas. BMC Cancer 18:270 (2018). PubMed: 29523123
    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

    Immunocytochemistry abreview for Anti-CD63 antibody [NK1/C3]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (cancer)
    Specification
    cancer
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°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 Jun 04 2010

    Question

    Regarding ab81436 (TAPA1).   I am not working on blood or cells. I am working on exosomes. CD81 (TAPA1) is a marker for exosomes, usually I will have exosomes in 100ul of PBS, I would like to ask since your catalog suggested using 100 ul blood and 20ul Ab, so I presume it will be same for exosomes in PBS. but I like to ask should I add 20 ul of this ab onto exosomes pallets alone or 20 ul of this ab into exosomes in 100ul PBS?   and what concentration exactly of ab81200 (PE Control) should I use since I do not have the concentration of ab81436. Would you provide or suggest an exact concentration please.  

    Read More

    Abcam community

    Verified customer

    Asked on Dec 07 2011

    Answer

    Thank you for contacting us. If you are not able to easily pellet the exosomes, I would indeed recommend to add 20 µl of the antibody solution to the exosomes in 100 µl PBS. You may need to increase the incubation time and or the volume of ab81436 added, in order to optimise the staining conditions for this particular setting. To our knowledge, this has never been done before, so I am sorry that we cannot give you more detailed information. However, I trust that you will succeed! Unfortunately, it is NOT possible to determine the EXACT concentration of ab81436: The concentration of the specific TAPA1 antibodies has not been determined before conjugation. Even if it had been, this concentration would be different from the concentration of the antibody after the conjugation procedure, as there is always a loss during the purification steps. Determining the protein concentration after the conjugation does not result in accurate measurements, because Phycoerythrin (PE) is a huge protein itself and thus would give false results, as my colleagues tried to explain earlier (in case you did not receive these emails, see below). We would therefore suggest to use the PE isotype control ab81200 in the same dilution than ab81436. For example, if you add 20 µl of ab81436 to 100 µl of your sample, than it is recommended to also use 20 µl of the isotype control ab81200 for 100 µl of sample. Also, I would like to draw your attention to the criteria to record information about the experimental overview and samples (including controls) set out by the flow cytometry community (e.g. http://www.flowcytometryuk.org/). These criteria are known as the Minimum Information about a Flow Cytometry Experiment (MIFlowCyt), and contain valuable information about isotype controls and FMO controls. I hope this information is helpful to you. Please do not hesitate to contact me if you need any further advice or information in this regard. Previously: I can confirm that the concentration of this antibody was not determined. Regrettably, it is not possible to correctly measure the antibody concentration of products with antibodies conjugated to proteins (such as PE). /If there are concentrations given for conjugated antibodies, it is usually the IgG concentration measured before the conjugation took place and therefore only an estimate./ Unfortunately, we do not currently have more information about the concentration of ab81436./ Antibody concentration is usually determined by measuring the protein concentration. This is not useful in this case since PE is also a protein.

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2011

    Question

    Dear Sir/Madam: Recently we purchased ab1318 (anti-CD63) & ab92726 (anti-CD9) antibodies. What is the antibody concentration in these tubes? We don’t need the high accuracy, just the ballpark would be fine.  Thank you,  

    Read More

    Abcam community

    Verified customer

    Asked on Nov 01 2011

    Answer

    Thank you for contacting us with your question about these antibodies. We don't have exact concentrations for either of these products, but ab1318 is around 100 ug/mL. Since Ab92726 is provided as unpurified tissue culture supernatant, it is not possible to measure the concentration, but supernatant is estimated to have a concentration around 1-3 mg/mL of total protein. I hope this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Nov 01 2011

    Question

    Please could you tell me the concentration of antibody or total quantity in the 500 microlitre sized ab1318 ? Many thanks

    Read More

    Abcam community

    Verified customer

    Asked on Oct 13 2004

    Answer

    Thank you for your enquiry. The concentration is :100 ug/ml (micrograms per ml). Please do not hesitate to contact us if you need further information.

    Read More

    Abcam Scientific Support

    Answered on Oct 14 2004

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