Plasma/Serum Exosome Purification Kit

Supporting Data

Figure 1. Comparing exosome yields from different purification methods. Intact exosomes were purified from 1 mL plasma using different purification methods. Intact exosomes were purified from 1 mL plasma using Norgen's Plasma/Serum Exosome Purification Mini Kit (Cat# 57400), Competitor S's kit and Competitor L's kit (from plasma). Exosomes purified using Norgen's kit were resuspended in 200 µL of Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that Norgen's kit isolated 55 nm exosomes with a recovery of 9.64 x 10¹³ particles/mL plasma. No impurities were found to be contaminating the exosomes purified using Norgen's Plasma/Serum Exosome Purification Mini Kit. Additionally, exosomes with a broader size range covering from 50 nm - 150 nm were purified from 1 mL plasma with a higher concentration compared to the other two methods.

Figure 2. NanoSight data showing intact exosomes purified from 1 mL and 10 mL plasma. Intact exosomes were purified from 1 mL plasma using Norgen's Plasma/Serum Exosome Purification Mini Kit (Cat. 57400) and from 10 mL plasma using Norgen's Plasma/Serum Exosome Purification Maxi Kit (Cat. 57600). Exosomes purified using Norgen's Mini kit were resuspended in 200 µL of Norgen's ExoR buffer whereas exosomes purified using Norgen's Maxi kit were resuspended in 600 µL Norgen's ExoR buffer, diluted 1:1,000 and visualized on the NanoSight LM10 instrument. The analysis shows that the purification of exosomes is linear as 4.04 x 10¹⁰ particles/mL was recovered from 1 mL plasma whereas 2.95 x 10¹¹ particles/mL was recovered from 10 mL plasma.

Figure 3. TEM images of small EVs obtained from the same non disease frozen platelet free plasma (ND-frozen PFP) sample. Sample isolated with three commercial kits; representatives of >10 images are shown (scale bar, 200 nm).

(Human myeloma cell- and plasma-derived extracellular vesicles contribute to functional regulation of stromal cells. Journal Proteomics. 2021. Reale, A., Carmichael, I., Xu, R., Mithraprabhu, S., Khong, T., Chen, M., Fang, H., Savvidou, I., Ramachandran, M., Bingham, N. and Simpson, R.J.)

Figure 4. Western blot analysis of plasma small EVs (psEV). Plasma small EVs (psEV) were isolated with the Norgen Biotek kit (#57400) and lysates of psEV derived from frozen Non-disease platelet free plasma (ND-PFP) are compared to lysates of psEV derived from fresh platelet free plasma (PFP) using EV markers (TSG101, CD81, ALIX, CD63) and platelet markers (ITGA2B and SELP). Cell free RNA BCT-Streck (sRNA collection tubes) and EDTA tubes are also compared (n= 3).

(Human myeloma cell- and plasma-derived extracellular vesicles contribute to functional regulation of stromal cells. Journal Proteomics. 2021. Reale, A., Carmichael, I., Xu, R., Mithraprabhu, S., Khong, T., Chen, M., Fang, H., Savvidou, I., Ramachandran, M., Bingham, N. and Simpson, R.J.)

Kit Specifications
Plasma/Serum Input (Cat. 57400)	50 μL - 1 mL
Plasma/Serum Input (Cat. 57500)	1 mL - 4 mL
Plasma/Serum Input (Cat. 57600)	4 mL - 10 mL
Size of Exosomes Purified	40 nm - 150 nm
Elution Volume	Variable depending on the plasma/serum 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. This kit is stable for 2 years after the date of shipment.

Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Component	Cat. 57400 (50 preps)	Cat. 57500 (25 preps)	Cat. 57600 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 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

Documentation

Plasma/Serum Exosome Purification Kit