ExoBrite™ WGA EV Staining Kits

Fluorescent WGA conjugates that are optimized for bright staining of extracellular vesicles for flow cytometry.

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ExoBrite™ WGA EV Staining Kits - Image 2

SEC-purified EVs were stained with ExoBrite™ WGA EV Stains. MCF-7 derived EVs were stained with ExoBrite™ 410/450, ExoBrite™ 490/515, or ExoBrite™ 560/585. CHO derived EVs were stained with ExoBrite™ 640/660. 410/450 was detected in the Pacific Blue channel, 490/515 was detected in the FITC channel, 560/585 was detected in the PE channel, and 640/660 was detected in the APC channel.

ExoBrite™ WGA EV Staining Kits - Image 3

SEC purified MCF-7-derived EVs stained with ExoBrite™ 410/450 WGA EV Stain (left) compared to the same stain in buffer (right). EVs were detected on a CytoFLEX LX flow cytometer in the Pacific Blue channel.

ExoBrite™ WGA EV Staining Kits - Image 4

SEC purified MCF-7-derived EVs stained with ExoBrite™ 490/515 WGA EV Stain (left) compared to the same stain in buffer (right). EVs were detected on a CytoFLEX LX flow cytometer in the FITC channel.

ExoBrite™ WGA EV Staining Kits - Image 5

SEC purified MCF-7-derived EVs stained with ExoBrite™ 560/585 WGA EV Stain (left) compared to the same stain in buffer (right). EVs were detected on a CytoFLEX LX flow cytometer in the PE channel.

ExoBrite™ WGA EV Staining Kits - Image 6

SEC purified CHO-derived EVs stained with ExoBrite™ 640/660 WGA EV Stain (left) compared to the same stain in buffer (right). EVs were detected on a CytoFLEX LX flow cytometer in the APC channel.

ExoBrite™ WGA EV Staining Kits - Image 7

ExoBrite™ 490/515 WGA EV Stain was used to stain either SEC-purified, Jurkat-derived EVs, or artificial liposomes. Robust staining was seen with the EVs, but the liposomes were not stained. The liposomes in this experiment were prepared from Presome® ACD-1, and were verified to be of similar size and concentration to the EVs using the lipophilic dye di-8-ANNEPS (data not shown). Analysis was performed on a CytoFLEX flow cytometer with SSC detected from the 405 nm laser, and ExoBrite™ detected in the FITC channel. Presome® is a registered trademark of NIPPON FINE CHEMICAL CO., LTD.

ExoBrite™ WGA EV Staining Kits - Image 8

SEC-purified MCF-7-derived EVs were stained first with ExoBrite™ 490/515 CD81 in 100 uL, followed by 1X ExoBrite™ 640/660 WGA EV Stain. EVs were detected on a CytoFLEX LX flow cytometer in the FITC and APC channels. When we gated on ExoBrite™ 640/660 WGA-positive particles, ~60% were also positive for CD81.

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Product Description

ExoBrite™ WGA EV Staining Kits were designed to overcome some of the challenges of EV detection, particularly in flow cytometry. ExoBrite™ WGA EV Stains bind to molecules in the EV membrane for bright, specific staining, with little background.

Features

Optimally formulated WGA conjugates for staining purified EVs
Broad compatibility, stained EVs isolated from all 9 sources tested
Designed for detection by flow cytometry
Bright signal and low background
Compatible with antibody co-staining
Available in 4 colors

Kit Components

ExoBrite™ WGA EV Stain
ExoBrite™ 1X PBS Solution

ExoBrite™ WGA EV Stains are uniquely formulated conjugates of wheat germ agglutinin (WGA), a carbohydrate-binding lectin with high affinity for N-acetylglucosamine moieties of glycoproteins. WGA conjugates are often used for labeling cell membranes as well as gram-positive bacteria. WGA conjugates have also been used to detect EVs due to the presence of glycoproteins on EV membranes.

ExoBrite™ WGA EV Stains were designed to overcome some of the challenges of detecting isolated EVs, particularly in flow cytometry. For example, tetraspanin antibodies commonly used to stain EVs can have varying signal and coverage depending on the EV source. Conversely, ExoBrite™ WGA EV Stains show bright staining of EVs derived from a broad range of sources. We tested EVs derived from 9 cell lines and ExoBrite™ WGA EV Stains showed strong staining for all of them. ExoBrite™ WGA EV Stains are less prone to aggregation than hydrophobic membrane dyes and do not bind non-specifically to polystyrene beads, allowing them to be used to stain bead-bound EVs.

EVs are often labeled with fluorescent antibodies targeting one or more of the tetraspanin proteins CD9, CD63, and CD81. ExoBrite™ WGA staining can be combined with antibody staining, for multi-parameter analysis.

Notes:

ExoBrite™ WGA EV Stains have been found to label EVs derived from all cell lines tested (see Validated EV Sources below), but may not stain EVs from every source.
In our testing, we have found that ExoBrite™ 490/515 dye may bind to streptavidin coated surfaces or beads if free biotin binding sites are not blocked. We recommend performing a biotin blocking step after binding your biotinylated capture antibody to streptavidin beads or surfaces when using ExoBrite™ 490/515 conjugates. Alternatively, consider using a different ExoBrite™ dye for staining EVs captured on streptavidin beads or surfaces.

ExoBrite™ WGA EV Staining Kits

Product	Ex/Em	Detection channels	Size	Catalog Number
ExoBrite™ 410/450 WGA EV Staining Kit	416/452 nm	Pacific Blue™	100 Labelings	30123-T
ExoBrite™ 410/450 WGA EV Staining Kit	416/452 nm	Pacific Blue™	500 Labelings	30123
ExoBrite™ 490/515 WGA EV Staining Kit	490/516 nm	FITC	100 Labelings	30124-T
ExoBrite™ 490/515 WGA EV Staining Kit	490/516 nm	FITC	500 Labelings	30124
ExoBrite™ 560/585 WGA EV Staining Kit	562/584 nm	PE	100 Labelings	30125-T
ExoBrite™ 560/585 WGA EV Staining Kit	562/584 nm	PE	500 Labelings	30125
ExoBrite™ 640/660 WGA EV Staining Kit	642/663 nm	APC	100 Labelings	30126-T
ExoBrite™ 640/660 WGA EV Staining Kit	642/663 nm	APC	500 Labelings	30126

Validated EV Sources for ExoBrite™ EV Surface Stains

EV Source	ExoBrite™ True EV Membrane Stains	ExoBrite™ CTB Stains	ExoBrite™ WGA Stains	ExoBrite™ Annexin Stains
A549 cells	Yes	Yes	Yes	Yes
CHO cells	Yes	No	Yes	Yes
hASC (human adipose stem cells)	ND	No¹	ND	ND
HEK293 cells	Yes	Yes¹	Yes	ND
HeLa cells	Yes	No	Yes	Yes
HUVEC (human umbilical vein endothelial cells)	ND	No¹	ND	ND
J774 cells	Yes	Yes	Yes	Yes
Jurkat cells	Yes	Yes	Yes	Yes
MCF-7 cells	Yes	Yes	Yes	Yes
Plasma	ND	No	ND	Yes
Raji cells	ND	Yes	Yes	Yes
RAW 264.7 cells	Yes	ND	ND	ND
Serum	ND	No	ND	Yes
Skeletal myoblasts	ND	Yes¹	ND	ND
THP-1 cells	Yes	ND	ND	ND
U2OS cells	Yes	No	Yes	Yes
U937 cells	Yes	No	Yes	Yes
NIH3T3 cells	Yes	ND	ND	ND
HepG2 cells	ND	ND	Yes	ND
Yeast (S. cerevisiae)	Yes	No	Yes	Yes

¹Customer-reported data
Value of “Yes” or “No” indicates coverage of EVs based on Biotium’s internal data or customer-reported data. Value of “ND” indicates no data.

Biotium also offers other validated ExoBrite™ reagents for flow cytometry, western blotting, or super-resolution imaging.

Learn about Biotium's new ExoBrite™ True EV Membrane Stains. These genuine lipophilic membrane dyes are designed for superior pan-EV labeling over other membrane dyes including PKH, DiO, DiI, and DiD. Biotium also offers ExoBrite™ CTB EV Stains (cholera toxin B conjugates) and ExoBrite™ Annexin EV Stains optimized for bright and sensitive staining of EVs. The ExoBrite™ EV Surface Stain Sampler Kit contains each of Biotium’s ExoBrite™ EV Surface Stains (CTB, WGA, and Annexin V) for assessing which stain offers the best coverage for the EV samples of interest. Biotium also offers ExoBrite™ Antibody Conjugates for optimal detection of CD9, CD63, and CD81 EV markers by flow cytometry and western blotting. For super-resolution imaging by STORM, learn about our ExoBrite™ STORM CTB EV Staining Kits available in four CF® Dyes validated for STORM.

Product Attributes

Size

100 labelings, 500 labelings

Dye

ExoBrite™ 410/450, ExoBrite™ 490/515, ExoBrite™ 560/585, ExoBrite™ 640/660

Colors

Blue, Green, Orange-red, Far-red

Documents, Protocols, SDS and COA

Documents

ExoBrite Stains and Antibodies Brochure

Protocols

PI-ExoBrite WGA EV Staining Kits

SDS

SDS ExoBrite WGA EV Staining Kits

COA

Request the Certificate of Analysis

FAQs

Exosome & EV Staining

ExoBrite™ Antibodies for western validation of immunomodulatory MSC EVs

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

Can other membrane stains besides ExoBrite™ True EV Membrane Stains be used to stain extracellular vesicles (EVs)?

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.

Which of Biotium's antibodies can be used to stain EVs?

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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