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  1. Link

    cd63-antibody-epr5702-ab134045.pdf

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Cardiovascular Blood Platelets
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD63 antibody [EPR5702] (ab134045)

  • Datasheet
  • SDS
Reviews (6) Submit a question References (158)

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Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
  • Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
  • Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
  • Anti-CD63 antibody [EPR5702] (ab134045)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5702] to CD63 - Late Endosome Marker
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-CD63 antibody [EPR5702]
    See all CD63 primary antibodies
  • Description

    Rabbit monoclonal [EPR5702] to CD63 - Late Endosome Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD63 aa 100-200 (extracellular). The exact sequence is proprietary.

  • Positive control

    • WB: Human platelet, HL-60 and A375 cell lysates and human melanoma tissue lysate. Brefeldin A treated wild-type HAP1 whole cell lysate. IHC-P: Human melanoma and tonsil tissues.
  • 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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5702
  • Isotype

    IgG
  • 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

  • Alternative Versions

    • Anti-CD63 antibody [EPR5702] - BSA and Azide free (ab215821)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HL-60 whole cell lysate (ab7914)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab134045 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)
1/1000 - 1/10000. Detects a band of approximately 30-65 kDa (predicted molecular weight: 26 kDa).
IHC-P (1)
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

Notes
WB
1/1000 - 1/10000. Detects a band of approximately 30-65 kDa (predicted molecular weight: 26 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

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

    • Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      All lanes : Anti-CD63 antibody [EPR5702] (ab134045) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : Brefeldin A treated wild-type HAP1 whole cell lysate
      Lane 3 : CD63 knockout HAP1 whole cell lysate
      Lane 4 : Brefeldin A treated CD63 knockout HAP1 whole cell lysate
      Lane 5 : HL60 whole cell lysate
      Lane 6 : Human platelets whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 26 kDa



      Lanes 1 - 6: Merged signal (red and green). Green - ab134045 observed at 26 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab134045 was shown to specifically react with CD63 in wild-type HAP1 Brefeldin A treated cells as signal was lost in HAP1 Brefeldin A treated CD63 knockout cells. Wild-type and CD63 knockout samples were subjected to SDS-PAGE. Ab134045 and ab9484 (Mouse anti-GAPDH 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      All lanes : Anti-CD63 antibody [EPR5702] (ab134045) at 1/5000 dilution (purified)

      Lane 1 : A375 whole cell lysate
      Lane 2 : HL-60 whole cell lysate
      Lane 3 : Human melanoma tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 26 kDa
      Observed band size: 30-65 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling CD63 with unpurified ab134045 at a dilution of 1/250.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [EPR5702] (ab134045)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue (negative control) labelling CD63 with purified ab134045 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      Western blot - Anti-CD63 antibody [EPR5702] (ab134045)
      All lanes : Anti-CD63 antibody [EPR5702] (ab134045) at 1/1000 dilution (unpurified)

      Lane 1 : Human platelet lysate
      Lane 2 : Human melanoma lysate
      Lane 3 : HL-60 lysate
      Lane 4 : A375 lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 26 kDa

    • Anti-CD63 antibody [EPR5702] (ab134045)
      Anti-CD63 antibody [EPR5702] (ab134045)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (158)

    Publishing research using ab134045? Please let us know so that we can cite the reference in this datasheet.

    ab134045 has been referenced in 158 publications.

    • Wei D  et al. RAB31 marks and controls an ESCRT-independent exosome pathway. Cell Res 31:157-177 (2021). PubMed: 32958903
    • Liu X  et al. Hypoxic bone marrow mesenchymal cell-extracellular vesicles containing miR-328-3p promote lung cancer progression via the NF2-mediated Hippo axis. J Cell Mol Med 25:96-109 (2021). PubMed: 33219752
    • Cai G  et al. Mesenchymal stem cell-derived exosome miR-542-3p suppresses inflammation and prevents cerebral infarction. Stem Cell Res Ther 12:2 (2021). PubMed: 33407827
    • Ridzuan N  et al. Human umbilical cord mesenchymal stem cell-derived extracellular vesicles ameliorate airway inflammation in a rat model of chronic obstructive pulmonary disease (COPD). Stem Cell Res Ther 12:54 (2021). PubMed: 33436065
    • Li H  et al. Knockdown of circ-FANCA alleviates LPS-induced HK2 cell injury via targeting miR-93-5p/OXSR1 axis in septic acute kidney injury. Diabetol Metab Syndr 13:7 (2021). PubMed: 33468219
    View all Publications for this product

    Customer reviews and Q&As

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

    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-CD63 antibody [EPR5702]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - other (Exosomes of melanoma cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Exosomes of melanoma cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 23 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD63 antibody [EPR5702]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Melanoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA
    Permeabilization
    No
    Specification
    Melanoma
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 12 2021

    Immunocytochemistry abreview for Anti-CD63 antibody [EPR5702]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (HaCaT)
    Permeabilization
    Yes - 0,2% Triton X100
    Specification
    HaCaT
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 22 2017

    ELISA abreview for Anti-CD63 antibody [EPR5702]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Cell (Exosomes from MCF-7 cells)
    Specification
    Exosomes from MCF-7 cells
    Blocking step
    glycine as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 37.5µg/mL · Temperature: 37°C
    Type
    Sandwich (Detection)
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 26 2016

    Flow Cytometry abreview for Anti-CD63 antibody [EPR5702]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (hFOB 1.19)
    Permeabilization
    No
    Specification
    hFOB 1.19
    Preparation
    Cell harvesting/tissue preparation method: Cel line from ATCC
    Sample buffer: PBS pH 7.2, BSA 0.5%, EDTA 2 mM
    Fixation
    --
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 26 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CD63 antibody [EPR5702]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Sample
    Human Cell (Blood film)
    Specification
    Blood film
    Permeabilization
    Yes - 0.1M PBS with 1% TritonX100
    Fixative
    Paraformaldehyde
    Read More

    DR. Ruma Raha-Chowdhury

    Verified customer

    Submitted Sep 26 2014

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