ExoGlowTM-NTA Fluorescent Labeling Kit

See only extracellular vesicles with NanoSight for more accurate particle size analysis and quantitation

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ExoGlow™-NTA Fluorescent Labeling Kit

10 Reactions
EXONTA100A-1
$375

Overview

Just see extracellular vesicles with NanoSight

Making a great exosome research tool even better, SBI has developed ExoGlowTM-NTA, a proprietary dye that enables fluorescent NanoSightTM analysis* of only the extracellular vesicles present in a heterogenous sample. The result is more accurate EV NanoSight data that excludes protein aggregates, membrane fractions, and other background particles to provide EV-specific particle size and concentration.

  • The only commercially available kit that specifically labels EVs for fluorescent NanoSight NTA quantitation
  • Delivers high signal-to-noise ratio with a proprietary dye that specifically binds EVs
  • Validated using common EV isolation methods including as ExoQuickTM, ultracentrifugation, and column-based methods (see Supporting Data, Figure 1)
  • Optimized for a fast protocol that takes only 45 minutes from sample isolation to analysis

*For fluorescent NTA analysis, the NanoSight NTA system (model LM10, NS300 or others) MUST be installed and properly calibrated with a 488nm laser (available separately). Please contact your local Malvern Instruments representative for additional information.

How It Works

Gain more accurate insight into your exosome sample

The ExoGlow-NTA Kit takes advantage of the fluorescence capabilities of the NanoSight instrument* with a proprietary fluorescent dye, which works by binding specifically and efficiently to the surface of intact vesicles. Membrane fragments, protein aggregates, and other background particles do not bind the ExoGlow-NTA dye, resulting in exclusion of these species from fluorescent NTA analysis (Figure 1). Thus, with the ExoGlow-NTA Kit, the data delivered by NTA more accurately represents the EV populations in your sample rather than all particles, as is typically reported by conventional (non-fluorescent) NTA.

ExoGlow-NTA dye only binds to the membranes of intact EVs

Figure 1. The ExoGlow-NTA dye only binds to membranes of intact EVs. Unlike conventional NTA, which collects data on all particles in a solution based on light scattering, the fluorescence mode of the NanoSight instrument selectively detects the labeled EVs and only the data from fluorescently-labeled particles is reported.

The ExoGlow-NTA Fluorescent Labeling Kit comes with three components: 1) Labeling dye 2) Internal Standards and 3) Reaction Buffer. Simply mix the dye with the reaction buffer, add 1-100 µg of EVs (or protein equivalent), incubate for 30 minutes, and you are ready for fluorescent NTA analysis. The provided Internal Standards are size-controlled synthetic liposomes that provide a positive control for NanoSight calibration as well as EV/exosome labeling efficiency using the ExoGlow-NTA Kit.

*For fluorescent NTA analysis, the NanoSight NTA system (model LM10, NS300 or others) MUST be installed and properly calibrated with a 488nm laser (available separately). Please contact your local Malvern Instruments representative for additional information.

Don’t have access to a NanoSight instrument equipped with the 488nm laser? SBI also offers fluorescent NTA as a service. Simply send us your samples, and we will isolate, label, and perform fluorescent NTA on your EVs. Email our Services department to request a quote now.

Supporting Data

Better NanoSight NTA data on exosomes using ExoGlow-NTA

SBI’s ExoGlow-NTA Fluorescent Labeling Kit delivers highly accurate EV quantitation in a quick and easy workflow. The proprietary dye offers highly efficient labeling (Figure 1), very low background (Figure 2), and is compatible with a wide range of EV isolation techniques (Figure 3).

ExoGlow-NTA-labeled liposome deliver NanoSight NTA data whether in light scattering or fluorescent mode.

Figure 1. ExoGlow-NTA-labeled liposomes deliver consistent NanoSight NTA data whether in light scattering or fluorescent mode. The high concordance of NTA and fluorescent NTA data collected from the ExoGlow-NTA Kit internal standards (ExoGlow-NTA-labeled synthetic liposomes) demonstrates the labeling efficiency of the ExoGlow-NTA Dye and accuracy of the fluorescent NTA method for characterizing EVs.

ExoGlow-NTA delivers undetectable background signal.

Figure 2. ExoGlow-NTA delivers undetectable background signal. When analyzing the ExoGlow-NTA dye alone in PBS, conventional NTA picks up background particles in the absence of EVs, while fluorescent NTA of the ExoGlow-NTA dye alone shows bias-free undetectable autofluoresence, based on (A) particle counts and (B) imaging.

ExoGlow-NTA demonstrates that conventional NTA overestimates EV concentration in samples irrespective of EV isolation method.

Figure 3. ExoGlow-NTA demonstrates that conventional NTA overestimates EV concentration in samples irrespective of EV isolation method. Representative data comparing conventional NTA and fluorescent NTA for EVs isolated using (A) ExoQuick (10 µg serum protein), (B) ultracentrifugation and wash (1 µg serum protein), or (C) column-based isolation (1 µg serum protein), shows just how much of the conventional NTA signal is due to non-EV particles.