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MigraPrep™︎ - Cultured-Cell Migrasome Production Kit (Crude Isolation)
PRC002
For migrasome production and crude isolation from cultured cells. The crude preparation can undergo further density-gradient or negative-selection purification.
Product manualThis kit is used for crude isolation of migrasomes from cultured cells. Refer to the manual for the full procedure and limitations.
The crude migrasome preparation can be further purified by conventional density-gradient separation with PRC003/PRC003S or by negative selection with PRC005/PRC005S.
Migratory adherent cell lines or primary cells are recommended. Migrasome production varies with the cells and must be evaluated experimentally. Before production, assessment of migrasome formation by in situ WGA staining and microscopy is recommended.
Components and storage
| Component | Qty. | Storage |
|---|---|---|
| Matrix solution J | PRC002: 1.5 mL; PRC002S: 300 µL | Ship at 4°C; store at -20°C on receipt. |
| Diluent K | PRC002: 50 mL; PRC002S: 10 mL | Ship at 4°C; store at 4°C on receipt. |
| Instruction manual | PRC002: 1 copy; PRC002S: 1 copy | Not applicable. |
PRODUCT FEATURES
Product advantages
Crude isolation of cultured-cell migrasomes
Provides a standardized preparation workflow from cell seeding and migrasome production to crude isolation by differential centrifugation.
Straightforward procedure
Requires routine cell culture and centrifugation at up to 18,000× g, without ultracentrifugation or magnetic-bead purification.
Compatible with multiple cultured cell types
Suitable for migratory adherent cell lines and primary cells; production conditions can be optimized for the characteristics of each cell type.
Compatible with multiple purification routes
The crude migrasome preparation can undergo conventional density-gradient purification or immunomagnetic negative-selection purification.
EXPERIMENTAL WORKFLOW
Workflow
FAQ
Frequently asked questions
Does PRC002 include further purification?
The kit produces a crude migrasome pellet. Further purification uses PRC003/PRC003S for conventional density-gradient separation or PRC005/PRC005S for negative selection.
Is the supplied diluent K used directly for the 1:300 mixture?
First prepare diluted K at a PBS:reagent K ratio of 9:1. Then prepare the pretreatment solution at a matrix solution J:diluted K ratio of 1:300.
How should cell density and culture time be selected?
Optimize these conditions for the cell source. A confluence of 20–40% at collection is recommended, with a lower density for faster-migrating cells; 12–16 h of production is generally recommended.
Which centrifugation steps retain the supernatant or pellet?
Retain the supernatant after 300×g for 5 min and after 2,500×g for 10 min. After 18,000×g for 60 min, discard the supernatant and retain the crude migrasome pellet. Perform all three spins at 4°C.
Which centrifuge rotors are recommended?
A swing-bucket rotor is recommended for low-speed centrifugation. A fixed-angle rotor with an angle greater than 45° is recommended for high-speed centrifugation.
How should the crude migrasome preparation be stored?
Select storage conditions according to downstream use. For analysis of contents, such as nucleic acid extraction, Western blotting or mass spectrometry, remove the supernatant after high-speed centrifugation and store the pellet at -80°C. Storage at 4°C is recommended when morphology, number, surface properties or activity are relevant. Storage at 4°C for up to one week has relatively little effect on morphology and number. Freezing at -20°C or -80°C causes aggregation and some rupture; a preliminary test is recommended if freezing is necessary.
Which reagents and equipment are supplied by the user?
Cell-culture reagents and consumables, including trypsin and culture dishes; sterile 1× PBS; low-binding 1.5 mL microcentrifuge tubes; 15 mL and 50 mL centrifuge tubes; low-binding pipette tips; a low-speed swing-bucket centrifuge; and a high-speed fixed-angle centrifuge.
Manual and COA
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