Exosome Isolation: Ultraclean EV PreparationUncompromising EV isolation delivers higher yields and cleaner preps
Important for intercellular communication in both normal physiology as well as disease states such as cancer, extracellular vesicle (EV) biology is a rapidly growing field. However, obtaining EVs for in vivo and ex vivo studies can be challenging. Ultracentrifugation has been considered the gold standard for exosome isolation, but the method is time-consuming, requires large sample volume inputs and access to specialized equipment.
Additionally, ultracentrifugation doesn’t isolate the cleanest exosome preparations it once was thought to produce. Other commercial kit are faster and easier than ultracentrifugation, but still include carryover protein that can cause over-estimation of EV amount and that can interfere with protein-sensitive studies such as mass spectrometry.
Now, with the newest generation of ExoQuick(-TC), ExoQuick(-TC) ULTRA, developed by our partner SBI you no longer have to make the trade-off between yield, purity, speed, a simple protocol, and price.
From as little as 250 uL of serum or plasma using the ExoQuick ULTRA kit (or as little as 5 mL of tissue culture media or other biofluid using the ExoQuick-TC ULTRA kit), you can isolate high-quality EVs for a wide range of downstream applications such as Western blot analysis, mass spectrometry, next generation sequencing, exosome labeling, and in vivo/ex vivo exosome delivery.
• Even cleaner—reduces carry-over of albumins by 75% and immunoglobulins by 40% more than other methods, including ultracentrifugation and competitors’ kits
• Higher yields—isolate more EVs per normalized input volume than ultracentrifugation and competitors’ kits
• Better biomarker detection—see what you’ve been missing when you increase the sensitivity of EV biomarker detection by up to 11-fold (CD9) and 8-fold (CD81)
• Fast—requires less than 20-minutes of hands-on time, including a convenient column-based format for clean-up
• Cost-effective—save money with each reaction compared to how much you’ll spend using a competitor’s kit
How to get to ULTRA-clean EV preps
The ExoQuick ULTRA protocol is quick and easy, requiring only 10-minutes of hands-on time:
Figure 1. ExoQuick ULTRA delivers high yields of clean exosomes. (A) A coomassie blue-stained protein gel comparing the protein content of exosome preps isolated using different methods shows only a few, defined protein bands in the ExoQuick ULTRA lane compared to the other methods. (B) Western blotting of the gel shows that the ExoQuick ULTRA prep contains the highest levels of exosome-specific markers CD9, CD81, and Hsp70 and the lowest levels of the carryover proteins albumin and IgGH. In contrast, the prep from Company Q appears to be primarily albumin, and even the sample prepared using ultracentrifugation contains considerably higher levels of both albumin and IgGH. Each lane was loaded with 7 ug of total protein as measured using a fluorometric Qubit protein assay.
Figure 2. Fluorescent nanoparticle tracking analysis (fNTA) demonstrates the high EV yields delivered by ExoQuick ULTRA compared to Ultracentrifugation. Comparison of different isolation methods on EV yields by both volume of input serum (per mL, A) and amount of input serum protein (per mg as measured by fluorometric Qubit protein assay, B). Particle number was measured using fNTA, a technique which specifically detects EVs.
Figure 3. EVs isolated using ExoQuick ULTRA display typical EV morphology. Transmission electron micrographs of EVs isolated from human serum using ExoQuick. The same sample is shown at two different magnifications. Multiple vesicles with typical EV morphology can be seen in each image.
|ExoQuick ULTRA EV Isolation Kit for Serum and Plasma||EQULTRA-20A-1-SBI||20 rxns||
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|ExoQuick-TC ULTRA EV Isolation Kit for Tissue Culture Media||EQULTRA-20TC-1-SBI||20 rxns||
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