Urine Exosome Purification Kit

Supporting Data

Figure 1. Intact exosomes purified from 5 mL using different purification methods. Intact exosomes were purified from 5 mL urine using Norgen's Urine Exosome Purification Midi Kit (Cat# 57800), Competitor S's kit, and Competitor L's kit. Exosomes purified using Norgen's kit were resuspended in 400 µL Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that Norgen's kit isolated 115 nm exosomes with a recovery of 8.36 x 10⁹ particles/mL urine samples. No impurities were found to be contaminating the exosomes purified using Norgen's Urine Exosome Purification Midi Kit. Additionally, Exosomes with a broader size range covering from 75nm - 250nm were purified from 5 mL urine with a higher concentration as compared to the other two methods.

Figure 2. Intact Exosomes were purified from 10 mL urine using Norgen's Urine Exosome Purification Midi Kit (Cat# 57800) and Ultracentrifugation. Exosomes purified using Norgen's kit and ultracentrifugation were resuspended in 400 µL of Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that Norgen's kit purified exosomes with sizes ranging from 65 nm to 195 nm, with a total recovery of 7.63 x 10⁸ particles/mL. No impurities were found to be contaminating the exosomes purified using Norgen's Urine Exosome Purification Midi Kit as opposed to the exosomes purified using ultracentrifugation, which purified exosomes with larger particle sizes ranging from 125 nm - 235 nm with a total recovery of 1.56 x 10⁸ particles/mL.

Kit Specifications
Urine Input (Cat. 57700)	250 μL - 1 mL
Urine Input (Cat. 57800)	2 mL - 10 mL
Urine Input (Cat. 57900)	11 mL - 30 mL
Size of Exosomes Purified	40 nm - 150 nm
Elution Volume	Variable depending on the urine input volume
Time to Complete 10 Purifications	15 - 30 minutes

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 57700 (50 preps)	Cat. 57800 (25 preps)	Cat. 57900 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Mini Filter Spin Columns inserted into 2 mL tubes	50	25	15
Product Insert	1	1	1