ExoBrite™ 490/515 WGA EV Staining Kit, 500 labelings
Referentie 30124
Formaat : 1kit
Merk : Biotium
ExoBrite™ WGA EV Staining Kits
Fluorescent WGA conjugates that are optimized for bright staining of extracellular vesicles for flow cytometry.
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.
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.
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.
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.
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.
HeLa EVs were stained with ExoBrite™ 490/515 WGA Stain with or without ExoBrite™ EV Stain Enhancer. The Enhancer reduces dye aggregates and increases the number of detected stained EVs, for improved signal-to-noise.
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.
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.
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.
The ExoBrite™ EV Stain Enhancer may be used to improve the signal-to-noise when staining with ExoBrite™ WGA EV Stains. The enhancer has shown to be beneficial for staining EVs with Annexin V, WGA, and other lectins. It is easy to use, simply add it directly to your staining reaction, and does not interfere with antibody staining.
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 |
| 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 |
ExoBrite™ EV Stains Comparison Guide
| ExoBrite™ EV Surface Stain | Pros | Cons |
|---|---|---|
| ExoBrite™ True EV Membrane Stains | • Near-complete staining of EVs in a sample • Broad compatibility with different EV sources • Validated for flow and fNTA | • Can't be used to stain bead-bound EVs • May have more aggregation than CTB & Annexin |
| ExoBrite™ Annexin EV Staining Kits | • Broad compatibility with different EV sources • Validated for flow and fNTA • Low background aggregates | • May not stain every EV in a sample • Doesn't work well on bead-bound EVs |
| ExoBrite™ WGA EV Staining Kits | • Broad compatibility with different EV sources • Can be used with bead-bound EVs | • May not stain every EV in a sample • Doesn't work well for fNTA |
| ExoBrite™ CTB EV Staining Kits | • Validated for flow and fNTA • Extremely low background • Can be used with bead-bound EVs | • May not stain every EV in a sample • Does not stain EVs from every source |
| ExoBrite™ Antibodies | • Highly specific for human tetraspanins CD9, CD63, CD81, and other EV markers • Validated for EV flow • Broad compatibility for different EV sources • Can be used with bead-bound EVs • Can be used for WB | • Depends on the expression level of the target protein on the EVs |
| ExoBrite™ EV Stain Enhancer | • Improves signal-to-noise by reducing or eliminating aggregates of certain EV stains • Validated with several different lectins and Annexin V • Does not interfere with antibody staining of EVs • Easy to use, just add directly to the staining reaction | • Not recommended for use with lipophilic EV stains |
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 | Yes |
| 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 | Yes | No | ND | Yes |
| Raji cells | ND | Yes | Yes | Yes |
| RAW 264.7 cells | Yes | Yes | Yes | Yes |
| Serum | Yes | 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 | Yes | Yes | Yes |
| HepG2 cells | Yes | No | Yes | Yes |
| Yeast (S. cerevisiae) | Yes | No | Yes | Yes |
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.
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