Great for Density Gradient Separation

September 20, 2017

University of California, Merced
Molecular and Cell Biology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

GE Healthcare Life Sciences

Product Name:

Ficoll-Paque PREMIUM

Catalog Number:

17-5442-02

We use this reagent to separate white blood cells from red blood cells using a ficoll density of 1.084. You can purchase different densities for different purposes. The separation is exact enough to isolate more specific populations of cell or reagents if needed. It is gentle enough to isolate live cells that can then be used for long term cell culture.

Experimental Design and Results Summary

Application

Blood Separation

Starting Material

Murine whole blood

Protocol Overview

Everything must be moved gently with very little perturbation to work optimally. We mix whole blood at a 1:1 ratio into PBS. Then layer 4ml of blood mixture onto 3ml of Ficoll. When pipetting ficoll, do not let is slosh onto the sides of the tube. When layering blood onto the surface of the ficoll, pipette slowly enough such that the blood do not break into the ficoll layer. Carefully move tubes containing blood and ficoll into a centrifuge and spin at 400g for 30 min. Allow centrifuge to come to a stop without applying any break (a break will cause the separated cells to mix). Isolate desired cell fraction.

Tips

Move slowly, any abrupt changes will cause undo mixing.

Results Summary

Separations appear as clean and distinct layers that can be separated by pipetting slowly.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Clean and clear distintions between sample layers.

The Bad

Easy to overmix sample into ficoll, making separation less clean.

The Bottom Line

You may have to optimize your hands but this will work to separate your samples by density gradient.

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