Exosomes Isolation Kit (Serum and Plasma )

IC-1006

Product Description

Exosomes are small vesicles(30¨C120 nm)containing RNA and protein that are secreted by various types of cells inculture,and found in abundance in body fluids includingblood,saliva,urine,and breast milk.Exosomes are thought to function as intercellular messengers,delivering their cargo of effector or signaling macromolecules between specificcells;however,their formation,the makeup of the cargo,and biological pathways in which they are involved remainincompletely understood.

The biological study of exosome function and traffickingrequires the isolation of intact exosomes,but the currentmethods used are tedious and difficult.The Total Exosome Isolation(from plasma)reagent provides a simple and reliable method of concentrating intact exosomes from human and mouse plasma samples.By tying up water molecules,the Total Exosome Isolation(from plasma) reagent forces less-soluble components(i.e.exosomes)out of solution,allowing them to be collected with a brief,low-speed centrifugation.
Prepare Sample

1. Remove the plasma sample from storage and place on ice. If the sample is frozen, thaw the sample in a 25¡ãC to 37¡ãC  water bath until it is completely liquid, and place on ice until needed.

2.Centrifuge the plasma sample at 2000 ¡Á g for 20 minutes at room temperature to remove cells and debris.

3.Transfer the supernatant containing the partially clarified plasma to a new tube without disturbing the pellet.

4.Centrifuge the new tube at 10,000 ¡Á g for 20 minutes at room temperature to remove debris.

5.Transfer the supernatant containing the clarified plasma to a new tube without disturbing the pellet, and place it on ice until ready to perform the isolation.

6.Proceed to ¡°Isolate Exosomes (with proteinase treatment),¡± or ¡°Isolate Exosomes (without proteinase treatment). ¡±

Isolate Exosomes (without proteinase treatment)

1.Transfer the required volume of clarified plasma to a new tube and add 0.5 volumes of 1X PBS.

2.Mix the sample thoroughly by vortexing.

3.Add 0.2 volume (i.e. Total volume = plasma + PBS) of the Exosome Precipitation Reagent (from plasma) to the sample.

4.Mix the plasma/reagent mixture well either by vortexing or inversion until the solution is homogenous.

Note: The solution should have a cloudy appearance.

5.Incubate the sample at room temperature for 10 minutes.

6.After incubation, centrifuge the sample at 10,000 ¡Á g for

5 minutes at room temperature.

7.Aspirate the supernatant by pipetting and discard.

Note: Exosomes are contained in a pellet at the bottom of the tube.

8.(Optional) Centrifuge the tube for 30 seconds at 10,000 ¡Á g to collect any residual reagent.

9.Discard any residual supernatant by careful aspiration with a pipet and proceed to ¡°Resuspend Exosomes.¡±

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