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ExoBrite™ CD63 Western Antibody

Validated antibody for optimal detection of EV marker CD63 in extracellular vesicle (EV) extracts by fluorescent western blot or chemiluminescence.

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ExoBrite™ CD63 Western Antibody
ExoBrite™ CD63 Western Antibody - Image 2Western detection of human CD63 in cell and exosome lysate using ExoBrite™ 680/700 CD63 Western Antibody, showing enrichment of CD63 in the exosome prep. Exosomes were isolated from MCF-7 cell conditioned medium by size exclusion column. The indicated amounts of lysates from MCF-7 cells and MCF-7-derived exosomes were run on an acrylamide gel and transferred to PVDF. The membrane was blocked with TrueBlack® Western Blocking Buffer, stained with 1X ExoBrite™ 680/700 CD63 Western Antibody, and imaged on a LI-COR Odyssey® infrared imaging system in the 700 channel. The protein ladder is Peacock™ Prestained Protein Marker, 1.5 uL per lane.
ExoBrite™ CD63 Western Antibody - Image 3Western blot showing CD63 enrichment in exosomes using ExoBrite™ Western Antibodies in MCF-7 cell and exosome lysates. Exosomes were isolated from MCF-7 cell conditioned medium by size exclusion column. The indicated amounts of lysates from MCF-7 cells and MCF-7-derived exosomes were run on an acrylamide gel and transferred to PVDF. The membrane was blocked with TrueBlack® Western Blocking Buffer, stained with 1X ExoBrite™ 680/700 CD63 Western Antibody and 1X ExoBrite™ 770/800 Calnexin Western Antibody, and imaged on a LI-COR Odyssey® infrared imaging system in the 700 and 800 channels. The protein ladder is Peacock™ Prestained Protein Marker, 1.5 uL loaded.
ExoBrite™ CD63 Western Antibody - Image 4Western blot showing CD9 and CD63 enrichment in exosomes using ExoBrite™ Western Antibodies in MCF-7 cell and exosome lysates. Exosomes were isolated from MCF-7 cell conditioned medium by size exclusion column. The indicated amounts of lysates from MCF-7 cells and MCF-7-derived exosomes were run on an acrylamide gel and transferred to PVDF. The membrane was blocked with TrueBlack® Western Blocking Buffer, stained with 1X ExoBrite™ 680/700 CD63 Western Antibody and 1X ExoBrite™ 770/800 CD9 Western Antibody, and imaged on a LI-COR Odyssey® infrared imaging system in the 700 and 800 channels. The protein ladder is Peacock™ Plus Prestained Protein Marker, 3 uL loaded.
ExoBrite™ CD63 Western Antibody - Image 5Western blot showing CD63 and CD81 enrichment in exosomes using ExoBrite™ Western Antibodies in MCF-7 cell and exosome lysates. Exosomes were isolated from MCF-7 cell conditioned medium by size exclusion column. The indicated amounts of lysates from MCF-7 cells and MCF-7-derived exosomes were run on an acrylamide gel and transferred to PVDF. The membrane was blocked with TrueBlack® Western Blocking Buffer, stained with 1X ExoBrite™ 680/700 CD81 Western Antibody and 1X ExoBrite™ 770/800 CD63 Western Antibody, and imaged on a LI-COR Odyssey® infrared imaging system in the 700 and 800 channels. The protein ladder is Peacock™ Plus Prestained Protein Marker, 1.5 uL loaded.

Product Description

ExoBrite™ CD63 Western Antibody is validated by Biotium for optimal detection of extracellular vesicle (EV) marker CD63 in EV extracts by fluorescent western blot. It is available conjugated to ExoBrite™ 680/700 or ExoBrite™ 770/800 near-infrared fluorescent dyes, which offer greater signal-to-noise than visible light fluorescent dyes for western blotting, as well as in an HRP-conjugated format for chemiluminescent detection.

  • Optimal detection of EV marker CD63 by fluorescent western blot or chemiluminescence
  • Validated for use with EV extracts
  • Bright signal and low background
  • Available in 2 near-infrared colors and HRP

EVs, including exosomes, are lipid-bound vesicles that are released from cells. EVs display specific surface proteins and can carry nucleic acids and other cargo, allowing them to transfer biological information between cells in different parts of the body. Therefore EVs are increasingly studied for their potential use in drug delivery and medical diagnostic applications. The most common proteins used as EV markers are CD9, CD63, and CD81, members of the tetraspanin family. Tetraspanins are plasma membrane proteins with many proposed functions, including activation and sorting of other membrane proteins. They are also thought to play a role in the targeting of proteins to multivesicular bodies (MVBs) and exosomes. These tetraspanins are broadly expressed on many cell types and can therefore be detected on many types of EVs, but their expression levels vary depending on the cell type of origin.

EV antibodies you can trust

Other commercially available antibodies for tetraspanin proteins CD9, CD63, and CD81 are generally not validated for isolated EVs and may require tedious optimization for your EV prep and staining protocol. ExoBrite™ Western Antibody Conjugates were validated to offer bright signal and low background of EV markers in EV extracts. ExoBrite™ Calnexin Western Antibody detects a protein of the endoplasmic reticulum that is not found in EVs. It is offered as a negative control to assess the purity of isolated EV extracts.

If you are using secondary antibodies for western detection, Biotium also offers unconjugated recombinant antibodies against CD9, CD63, and CD81. ExoBrite™ Flow Antibody Conjugates are also available for optimal detection of CD9, CD63, and CD81 EV markers by flow cytometry.

For general EV staining, Biotium offers ExoBrite™ stains conjugated to cholera toxin B (CTB), wheat germ agglutinin (WGA), and Annexin V. These stains are specially formulated for bright and specific detection of isolated EVs by flow cytometry. These ExoBrite™ stains may also be combined with antibody staining, for multi-parameter analysis.

Biotium also provides ExoBrite™ STORM Antibodies against CD9, CD63, and CD81, as well as ExoBrite™ STORM CTB EV Stains that use CF® Dyes engineered specifically for high-performance super-resolution imaging by STORM.

ExoBrite™ Western Antibody Conjugates

AntibodyEx/EmConc.SizeCatalog No.
ExoBrite™ 680/700
CD9 Western Antibody		681/698 nm100 ug/mL25 testsP003-680-250
100 testsP003-680-1000
ExoBrite™ 770/800
CD9 Western Antibody		770/797 nm100 ug/mL25 testsP003-770-250
100 testsP003-770-1000
ExoBrite™ HRP
CD9 Western Antibody		N/A100 ug/mL50 testsP003-HRP-500UL
ExoBrite™ 680/700
CD63 Western Antibody		681/698 nm100 ug/mL25 testsP004-680-250
100 testsP004-680-1000
ExoBrite™ 770/800
CD63 Western Antibody		770/797 nm100 ug/mL25 testsP004-770-250
100 testsP004-770-1000
ExoBrite™ HRP
CD63 Western Antibody		N/A100 ug/mL50 testsP004-HRP-500UL
ExoBrite™ 680/700
CD81 Western Antibody		681/698 nm100 ug/mL25 testsP006-680-250
100 testsP006-680-1000
ExoBrite™ 770/800
CD81 Western Antibody		770/797 nm100 ug/mL25 testsP006-770-250
100 testsP006-770-1000
ExoBrite™ HRP
CD81 Western Antibody		N/A100 ug/mL50 testsP006-HRP-500UL
ExoBrite™ 770/800
Calnexin Western Antibody		770/797 nm100 ug/mL25 testsP007-770-250
100 testsP007-770-1000

Product Attributes

Antibody number
#P004
SwissProt
P08962
Antibody type
ExoBrite™, Primary
Clonality
Monoclonal
Host species
Mouse
Isotype
IgG1, kappa
Antibody reactivity (target)
CD63
Synonyms
gp55; granulophysin; Lysosomal-associated membrane protein 3 (LAMP-3); Mast cell antigen AD1; melanoma 1 antigen; Melanoma-associated antigen MLA1; Melanoma-associated antigen ME491; MLA1; NGA; Ocular melanoma-associated antigen; OMA81H; PTLGP40; Tetraspanin-30; TSPAN30
Species reactivity
Baboon, Cynomolgus monkey, Human, Non-human primates
Human gene symbol
CD63
Entrez gene ID
967
Unigene
445570
Molecular weight of antigen
26 kDa (core protein); 30-60 kDa (glycosylated)
Antibody target cellular localization
Exosomes/EVs, Lysosomes, Plasma membrane, Membrane/vesicular, Multivesicular bodies
Cell/tissue expression
Exosomes, Platelets, Granulocytes, Lymphocytes, Monocytes/macrophages
Verified antibody applications
WB (verified)
Shipping condition
Room temperature
Positive control
MCF-7 cells, MCF-7 derived exosomes
Antibody application notes
Optimal concentration to be determined by end-user, Recommended amount for western blot: 100 ng/mL = 1:1000 dilution
Antibody research areas
Exosomes/EVs
Antibody/conjugate formulation
Fluorescent conjugates: proprietary buffer containing 0.05% sodium azide, HRP conjugates: PBS/50% glycerol/2 mg/mL rBSA
Shelf life
Guaranteed for at least 24 months from date of receipt when stored as recommended
Storage Conditions
Store at 2 to 8 °C, Protect fluorescent conjugates from light
Regulatory status
For research use only (RUO)
Product origin
Product may contain either bovine serum albumin (BSA) from bovine serum (Bos taurus), or recombinant BSA produced in Chinese hamster ovary cells. Inquire for the specific lot.

Documents, Protocols, SDS and COA

FAQs

Exosome & EV Staining

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) are emerging as powerful, cell-free immunomodulatory therapies for inflammatory diseases such as COVID-19. However, because the mechanism is poorly understood, optimizing EV-based therapies remains challenging.

In a 2025 Springer Nature study, Infante et al. investigated how COVID-19 patient serum reshapes the transcriptome and paracrine activity of Wharton’s jelly–derived MSC stem cells (WJ-MSCs). WJ-MCSs exposed to serum from hospitalized COVID patients showed downregulation of NEAT1 and MALAT1, two pro-inflammatory two long noncoding RNAs (lncRNAs). Furthermore, the researchers found that EVs derived from the treated cells had enhanced immunosuppressive activity when administered to T-cells.

The researchers isolated EVs from WJ-MSC cells after NEAT1 and/or MALAT1 knockdown, and tested whether there was an effect on T-cell proliferation. A Western blot of EVs derived from control and lncRNA-knockdown MSCs were probed with ExoBrite™ 680/700 CD81 Western Antibody. ExoBrite™ 770/800 Calnexin Western Antibody was also used as an endoplasmic reticulum marker to assess cellular contamination.

EV enriched samples in control, NEAT1 knockdown, MALAT1 knockdown, and NEAT1/MALAT1-double knockdown were confirmed by bright CD81 detection and the absence of Calnexin. They found that the MALAT1 knockdown EVs were found to have an inhibitory effect on T-cell proliferation. These results illustrate the importance of EV characterization using tools like Biotium’s ExoBrite™ antibodies in translational EV research.

Isolation and characterization of EVs from various lncRNA knock-down WJ-MSCs. Western blot analysis using ExoBrite™ 680/700 CD81 and ExoBrite™ 770/800 Calnexin in EV and MSC lysates. Asterisk (*) indicates reduced conditions used in the MSCs lysate. Modified from Infante et. al. Reproduced under CC BY 4.0.

Learn more about Biotium’s many stains and antibodies for EV research, including ExoBrite™ CD9/CD63/CD81 Antibody Cocktails for flexible and bright multiplexing detection by flow cytometry. Biotium also offers ExoBrite™ stains for pan-EV labeling, optimized fluorescent conjugates of CTB, WGA, and Annexin V for EV detection, ExoBrite™ antibodies for STORM imaging, and more.

Full Citation:

Infante, A., Cabodevilla, L., Gener, B. et al. Modulation of NEAT1 and MALAT1 expression in WJ-MSCs by Covid-19 serum: a foundation for EVs-mediated therapy. Respir Res 26, 313 (2025). https://doi.org/10.1186/s12931-025-03394-4

While early studies of EVs attempted to use first-generation membrane dyes like DiI or PKH to stain EVs, more recently this class of dyes has been found to be largely unsuitable for EV staining due to their high degree of aggregation. Dye aggregation not only generates nonspecific particles that are indistinguishable from EVs in flow cytometry, but also results in poor EV labeling efficiency. Biotium developed the ExoBrite™  True EV Membrane Stains in response to our customers’ difficulties with using traditional membrane dyes to stain EVs. See our Literature Digest for more information.

We strongly recommend our ExoBrite™ Flow Antibody Conjugates for staining both purified or bead-bound EVs. The antibodies are validated and optimized to offer bright signal and low background. They are available against human or mouse CD9, CD63, and CD81 tetraspanin proteins.

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