SBI’s ExoGlow-Membrane EV Labeling Kit (Cat # EXOGM600A-1) is the latest generation of fluorescent labeling reagent for robust and specific labeling of EV membranes. Based on a highly specific membrane sensor, the reagent specifically labels intact EVs for fluorescence-based detection, while displaying minimal background in an express.
As a result, the kit can be used for most applications that require the display of tagged electric vehicles for tracking studies. With very low intrinsic background levels, the tint is well suited for demanding EV labeling studies that require high specificity and signal-to-noise ratios.
The ExoGlow-Membrane Kit is shipped on ice and should be stored at + 4 ° C. Properly stored kits are stable for 12
months from the date of receipt.
- The size of the reaction is based on the use of 50-100 µg of exosomes.
- The excitation and emission values of the ExoGlow membrane in the bound state are 465 nm and 635 nm, respectively.
- We recommend mixing Reaction Buffer well before use.
- Protect the labeling dye from light.
- We recommend removing free dye before applying labeled exosomes to cells with PD MiniTrap or PD SpinTrap G-25 Buffer Exchange Column (GE, Cat # 28-9180-07 or 28-9180-04).
- Due to the nature of the sensor tint and its broad emission spectrum, the signal can also be detected in the green channel. Therefore, we do not recommend this product for multiplexing experiments.
- In 12 µl of Reaction Buffer, add 2 µl of Labeling Dye and mix well until the dye dissolves completely to make a labeling reaction buffer.
- Add 50-100 µg of exosomes to the labeling reaction buffer from step 1.
- Mix the sample well and incubate for 30 minutes at RT. It is not necessary to rotate the tubes during the incubation period. Protect tubes from light.
- Remove any unlabeled dye. We recommend the PD MiniTrap or PD SpinTrap G-25 Buffer Exchange Column by GE (not included). A single 1 minute spin at the recommended speed will be sufficient to remove the dye.
- Alternative protocol for free dye removal. If you have a high background after PD SpinTrap G-25, the reprecipitation protocol with ExoQuick-TC is a better option:
- Add 35 μL of ExoQuick-TC to 100 μL of Sample and incubate at 4o C from 30 min to overnight.
- Spin the Eppendorf tube at 10,000 rpm for 10 minutes.
- Gently aspirate the supernatant from the corner of the tube.
- Resuspend the EV pellet in PBS and continue with subsequent applications.
- Labeled exosomes can be added to cells or used for downstream applications