MagCapture(TM) Exosome Isolation Kit PS

When exosomes and other large EVs (microvesicles) are needed, prepare 1,200 × g supernatant^※1 as a sample. Additionally, when highly purified exosomes are needed, prepare 10,000 × g supernatant^※2 as a sample. This protocol for sample preparation is set for cell culture medium, serum, and plasma. When other body fluids are used, please examine the appropriate preprocessing protocol by referring to the protocol for serum and plasma.

Cell Culture Medium

Serum and Heparin Plasma

EDTA Plasma

Protocol for buffer exchange (ultrafiltration)

Perform buffer exchange of 1 mL centrifuged EDTA plasma sample with 50 mL of TBS buffer.

1. Add 19 mL of TBS to Vivaspin20 (100K).
2. Add the 1 mL of centrifuged EDTA plasma sup. to the Vivaspin20 (100K) of 1. and mix (solution A).
3. Centrifuge solution A at 4°C. 　（Refer to centrifugal condition in the manufacturer’s instruction manual.）
4. Add 10 mL of TBS to solution A when the upper liquid volume is dropped (solution B).
5. Centrifuge solution B at 4°C.
6. Add 10 mL of TBS to solution B when the upper liquid volume is dropped (solution C).
7. Centrifuge solution C at 4°C.
8. Add 11 mL of TBS to solution C when the upper liquid volume is dropped.
9. Centrifuge solution C at 4°C until the volume becomes under 1 mL.
10. Proceed to “Isolation step”.

※1 When exosomes and large EVs are needed, use 1,200 × g sup. fraction as sample.
※2 When exosomes are needed, use 10,000 × g sup. fraction as sample.
※3 When Large EVs are needed, use ppt of Large EVs obtained by centrifugation at 10,000 x g as sample after suspending it with TBS.
※4 The concentration step is an option when using a large volume (~ 50 mL) of cell culture supernatant as a sample for purification. However, since recovery efficiency improves, please perform it as much as possible.

Note : a volume of large or small sample
■In the case of large volume
The concentration of the sample is recommended when using a large volume (~50 mL) of cell culture supernatant as a sample for purification. Since recovery efficiency improves, please perform it as much as possible.

■In the case of small volume
Add the appropriate volume of TBS into samples to reach the volume of 0.5 mL to obtain the better mixture of the Exosome Capture-immobilized beads with the sample. (Example: 100-200 μL → 500 μL)

"This kit was the most successful exosome isolation method I have ever tried."

Biopharmaceutical company　|　Sample: Mesenchymal stem cells

"Because of the low back, the glycan analysis was successful. This kit provided the basis for determining exosomes as one of the theme in our laboratory."

University-Medical school　|　Sample: Human urine

"We have discovered a disease marker protein from serum-derived exosomes isolated with this kit."

University-Medical school　|　Sample: Human serum

"Exosomes are very pure with this kit compared to other methods of isolating exosomes depending on a sample."

Cancer research organization　|　Sample: Human plasma / serum

"Analysis of the exosomes isolated with this kit revealed candidate proteins for disease markers."

University-Medical school　|　Sample: Human urine