Overview

Move quickly with SBI’s fastest exosome quantitation method

With the EXOCET Exosome Quantitation Kit, you can find out just how many exosomes you’ve isolated in as little as 20 minutes. This antibody-free, colorimetric assay (OD405) measures the activity of Acetyl-CoA Acetylcholinesterase (AChE), an enzyme shown to be highly enriched in exosomes^{1, 2} (using mass spectrometry and Western blotting, SBI has verified the presence of AChE in exosomes from serum, stem cells, cancer cell lines, and MSCs—contact tech@systembio.com to learn more).

EXOCET is a highly quantitative assay and the standards included with each kit have been calibrated using NanoSight analysis, enabling calculation of the number of exosomes in your samples. The EXOCET Kit contains all of the necessary assay reagents and an optical 96-well plate to perform 96 reactions.

• Fast—complete in 20 minutes
• Antibody-free—quantitation is based on the activity of AChE, an enzyme enriched in most exosomes
• Quantitative—included calibration standards enable calculation of the number of exosome particles
• Flexible—works with all mammalian species tested (human, mouse, rat)
• Fully compatible—works with most exosome isolation methods including the ExoQuick family of reagents, ultracentrifugation, immunoaffinity purification, and chromatography

Choose the exosome quantitation method that’s best for your studies

REFERENCES

1. Savina A, et al. The exosome pathway in K562 cells is regulated by Rab11. J Cell Sci. 2002 Jun 15. 115(Pt 12):2505-15. PMID: 12045221.
2. Gupta S, and Knowlton AA. HSP60 trafficking in adult cardiac myocytes: role of the exosomal pathway. Am J Physiol Heart Circ Physiol. 2007 Jun. 292(6):H3052-6. PMID: 17307989.

How It Works

Exosome quantitation with EXOCET is quick and easy

1. Isolate exosomes (ExoQuick, ExoQuick-TC, ExoQuick PLUS, and ExoQuick-TC PLUS are all excellent methods)
2. Lyse exosomes with the included Exosome Lysis Buffer
3. Measure AChE activity—add the included buffer and incubate for 10–20 minutes. Readout is at 405 nm

Supporting Data

The fast, highly quantitative EXOCET exosome quantitation assay

Exosome quantitation with EXOCET takes as little as 20 minutes and 50 µg (protein equivalent) of sample. Exosomes were isolated from either human serum (0.5 mL) or from MDA-MB-231 culture media (10 mL) using standard ExoQuick and ExoQuick-TC reagents. Exosome pellets were resuspended in PBS and total protein concentration measured using a BCA assay (~10⁸ exosomes, 2 µg/µL). Exosomes were lysed using the EXOCET gentle lysis solution to preserve the enzymatic activity of the exosomal AChE enzyme. The standard curve was generated using known numbers of exosomes (as measured by NanoSight) and calibrated with a recombinant AChE enzyme standard solution provided in the kit.

Resources

Citations

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• Lee, JY, et al. (2016) Circulating exosomes from patients with systemic lupus erythematosus induce an proinflammatory immune response. Arthritis Res. Ther..2016 Nov 16; 18(1):264. PM ID:27852323
• Rossaint, J, et al. (2016) Directed transport of neutrophil-derived extracellular vesicles enables platelet-mediated innate immune response. Nat Commun.2016 Nov 15; 7:13464. PM ID:27845343
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