ExoBrite™ 490/515 WGA EV Staining Kit, 500 labelings

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.

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:

1. 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.
2. 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
			500 Labelings	30123
ExoBrite™ 490/515 WGA EV Staining Kit	490/516 nm	FITC	100 Labelings	30124-T
			500 Labelings	30124
ExoBrite™ 560/585 WGA EV Staining Kit	562/584 nm	PE	100 Labelings	30125-T
			500 Labelings	30125
ExoBrite™ 640/660 WGA EV Staining Kit	642/663 nm	APC	100 Labelings	30126-T
			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
HEK293T cells	ND	Yes1	ND	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	Yes	ND	ND	ND
Serum	ND	No	ND	Yes
Skeletal myoblasts	ND	Yes1	ND	ND
THP-1	Yes	ND	ND	ND
U2OS cells	Yes	No	Yes	Yes
U937 cells	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.