Density Gradient with Ficoll-Paque

December 13, 2018

BlueskyReddit
Johns Hopkins School of Medicine
Oncology
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

GE Healthcare Life Sciences

Product Name:

Ficoll-Paque PLUS

Catalog Number:

17144002

We use this reagent to isolate murine T lymphocytes after harvesting the spleen. This density gradient is extremely easy after practice and the yield is always clean and high.

Experimental Design and Results Summary

Application

T cell separation

Starting Material

Murine T cells

Protocol Overview

We bring up the T cells in RPMI and underlay with the Ficoll for easy separation of live and dead cells. This is especially important if we wish to use for adoptive transfer cells. After spinning for 30 minutes, there are three layers with the live cells in the middle layer. Simply extract the cloudy layer with a pipette.

Tips

Make sure reagent is in room temperature before proceeding.

Results Summary

The separation is always clear and easy to take out. The yield is always high with an average of 90% live cells.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use and great separation

The Bad

Reagent must be at room temperature otherwise the separation will be very convoluted

The Bottom Line

It is extremely easy to use without having to create your own density gradient. It is simply taken out from 4°C and rested until it reaches room temperature before using.

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