ExoBrite™ True EV Membrane Stains

Lipophilic membrane dyes designed specifically for EV research and the best choice for general pan-EV staining. The dyes are validated to offer higher (near-complete) coverage of EVs with lower aggregation than other membrane dyes like PKH, DiO, and DiI.

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ExoBrite™ True EV Membrane Stains - Image 2

ExoBrite™ 400/460 True EV Membrane Stain was used to stain SEC EVs isolated from MCF-7 cells. The samples were run on a CytoFLEX LX flow cytometer in the Pacific Blue channel. The gated population shows that stained EVs are easily distinguished from background, compared to the dye in buffer alone (right panel).

ExoBrite™ True EV Membrane Stains - Image 3

ExoBrite™ 515/540 True EV Membrane Stain was used to stain SEC EVs isolated from HeLa cells (left panel) or THP-1 cells (middle panel). The samples were run on a CytoFLEX LX flow cytmoeter in the FITC channel. The gated population shows that stained EVs are easily distinguished from background, compared to the dye in buffer alone (right panel).

ExoBrite™ True EV Membrane Stains - Image 4

ExoBrite™ 555/575 True EV Membrane Stain was used to stain SEC EVs isolated from HeLa cells (left panel) or THP-1 cells (middle panel). The samples were run on a CytoFLEX LX flow cytometer in the Y585 (PE) channel. The gated population shows that stained EVs are easily distinguished from background, compared to the dye in buffer alone (right panel).

ExoBrite™ True EV Membrane Stains - Image 5

ExoBrite™ 645/675 True EV Membrane Stain was used to stain SEC EVs isolated from MCF-7 cells (left panel). The samples were run on a CytoFLEX LX flow cytometer in the R660 (APC) channel. The gated population shows that stained EVs are easily distinguished from background, compared to the dye in buffer alone (right panel).

ExoBrite™ True EV Membrane Stains - Image 6

SEC purified MCF-7-derived EVs were stained with three green membrane dyes: ExoBrite™ 515/540 True EV Membrane Stain (left panel), PKH67 (middle panel), and Neuro-DiO (right panel). Of the three, only the ExoBrite™ True EV Membrane Stain is able to differentiate EVs from background particles in flow cytometry. The samples were run on a CytoFLEX LX flow cytometer in the FITC channel.

ExoBrite™ True EV Membrane Stains - Image 7

SEC purified MCF-7-derived EVs were stained with two orange membrane dyes: ExoBrite™ 555/575 True EV Membrane Stain (left panel), and DiI (right panel). Only the ExoBrite™ True EV Membrane Stain is able to differentiate EVs from background particles in flow cytometry. The samples were run on a CytoFLEX LX flow cytometer in the Y585 (PE) channel.

SEC purified MCF-7-derived EVs were stained with three green membrane dyes: ExoBrite™ 515/540 True EV Membrane Stain, PKH67, and Neuro-DiO. The stained EVs were diluted and run on a ZetaView particle analyzer. Total EV number was measured in scatter mode, and fluorescently-stained EV number was measured with 488 nm excitation and 500 nm filter. The percentage of particles stained with the dye is shown in the plot. ExoBrite™ 515/540 True EV Membrane Stain showed nearly complete coverage of EVs, much higher than PKH67 or Neuro-DiO.

SEC purified MCF-7-derived EVs were stained with two orange membrane dyes: ExoBrite™ 555/575 True EV Membrane Stain, and DiI. The stained EVs were diluted and run on a ZetaView particle analyzer. Total EV number was measured in scatter mode, and fluorescently-stained EV number was measured with 520 nm excitation and 550 nm filter. The percentage of particles stained with the dye is shown in the plot. ExoBrite™ 555/575 True EV Membrane Stain showed nearly complete coverage of EVs, much higher than DiI.

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Product Description

Elevate your EV research with ExoBrite™ True EV Membrane Stains. Offering unparalleled coverage and solubility, ExoBrite™ True EV Membrane Stains are genuine lipophilic membrane dyes that set a new standard in pan-EV labeling over classic dyes like PKH or DiO/DiI/DiD for EV staining.

Features

Lipophilic membrane dyes designed for pan-EV labeling
Superior alternative to common membrane dyes like PKH, DiO, DiI, DiD
Near-complete coverage of EVs in a sample
Broad compatibility with EVs isolated from different sources
Compatible with antibody co-staining
Available in 4 colors for Pacific Blue®, FITC, PE, and APC channels

Detect EVs, not dye aggregates

Membrane dyes like PKH, DiO, and DiI, while common tools for labeling EVs, can pose significant challenges when used for EV staining. For example, membrane dyes often have poor solubility and thus form aggregates that can be confused with EVs. ExoBrite™ True EV Membrane Stains were designed specifically to address the issues of membrane dye aggregation while remaining a true lipophilic membrane stain for pan-EV labeling.

ExoBrite™ True EV Membrane Stains are the best choice for general pan-EV staining. They demonstrate higher (near-complete) coverage of EVs over competitor membrane dyes in fluorescence nanoparticle tracking analysis (fNTA). The dyes also offer clear differentiation of EVs from non-specific particles during flow detection. ExoBrite™ True EV Membrane Stains labeled all EV types tested to date (see Validated EV Sources below).

Note: We do not recommend using ExoBrite™ True EV Membrane Stains to stain bead-bound EVs. For bead-bound EVs we recommend using ExoBrite™ CTB EV Stains or ExoBrite™ WGA EV Stains.

Significantly Greater Coverage of EVs Over Other Membrane Dyes Such as PKH, DiO, and DiI

SEC purified MCF-7-derived EVs were stained with three green membrane dyes: ExoBrite™ 515/540 True EV Membrane Stain, PKH67, and Neuro-DiO. The stained EVs were diluted and run on a ZetaView particle analyzer. Total EV number was measured in scatter mode, and fluorescently-stained EV number was measured with 488 nm excitation and 500 nm filter. The percentage of particles stained with the dye is shown in the plot. ExoBrite™ 515/540 True EV Membrane Stain showed nearly complete coverage of EVs, much higher than PKH67 or Neuro-DiO.

SEC purified MCF-7-derived EVs were stained with two orange membrane dyes: ExoBrite™ 555/575 True EV Membrane Stain, and DiI. The stained EVs were diluted and run on a ZetaView particle analyzer. Total EV number was measured in scatter mode, and fluorescently-stained EV number was measured with 520 nm excitation and 550 nm filter. The percentage of particles stained with the dye is shown in the plot. ExoBrite™ 555/575 True EV Membrane Stain showed nearly complete coverage of EVs, much higher than DiI.

Also view ExoBrite™ CTB, WGA, and Annexin EV Surface Stains

ExoBrite™ EV Surface Stains are fluorescent conjugates of probes for labeling EV membrane surface targets. The stains were designed to offer lower aggregation than competitor membrane dyes and are suitable for detecting bead-bound EVs, unlike ExoBrite™ True EV Membrane Stains and other hydrophobic membrane dyes. ExoBrite™ EV Surface Stains are available as cholera toxin subunit B (CTB), wheat germ agglutinin (WGA), or Annexin V conjugates. Biotium also offers a convenient ExoBrite™ EV Surface Stain Sampler Kit that includes each ExoBrite™ EV Surface Stains (CTB, WGA, and Annexin V) for assessing which stain offers the best coverage for the EV samples of interest.

Note: ExoBrite™ EV Stains have been found to label EVs derived from several tested cell lines (see Validated EV Sources below), but may not stain EVs from every source.

ExoBrite™ True EV Membrane Stains

Product	Ex/Em	Detection Channels	Size	Catalog Number
ExoBrite™ 400/460 True EV Membrane Stain	402/460 nm	Pacific Blue®	100 Labelings	30136-T
ExoBrite™ 400/460 True EV Membrane Stain	402/460 nm	Pacific Blue®	500 Labelings	30136
ExoBrite™ 515/540 True EV Membrane Stain	515/542 nm	FITC	100 Labelings	30129-T
ExoBrite™ 515/540 True EV Membrane Stain	515/542 nm	FITC	500 Labelings	30129
ExoBrite™ 555/575 True EV Membrane Stain	556/576 nm	PE	100 Labelings	30130-T
ExoBrite™ 555/575 True EV Membrane Stain	556/576 nm	PE	500 Labelings	30130
ExoBrite™ 645/675 True EV Membrane Stain	644/671 nm	APC	100 Labelings	30137-T
ExoBrite™ 645/675 True EV Membrane Stain	644/671 nm	APC	500 Labelings	30137

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 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.

Pacific Blue is a registered trademark of Thermo Fisher Scientific.

Product Attributes

Colors

Blue, Green, Red, Far-red

Detection method/readout

fNTA, Flow cytometry

Documents, Protocols, SDS and COA

Documents

Flow-Cytometry-Brochure

ExoBrite Stains and Antibodies Brochure

Protocols

PI-ExoBrite True Membrane EV Stains

SDS

SDS 30129-T, 30129, 30130-T, 30130, 30136-T, 30136, 30137-T, 30137_ExoBrite™ True EV Membrane Stains_US_en

SDS 30129-T, 30129, 30130-T, 30130, 30136-T, 30136, 30137-T, 30137_ExoBrite™ True EV Membrane Stains_BE_en

SDS 30129-T, 30129, 30130-T, 30130, 30136-T, 30136, 30137-T, 30137_ExoBrite™ True EV Membrane Stains_CA_en

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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