Easy Way to Isolate Polymorphonuclear Cells from Bone Marrow

February 09, 2016

BlueskyReddit
University of California, San Diego
Department of Pediatrics
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Mitenyi MACS

Product Name:

Neutrophil Isolation Kit, mouse

Catalog Number:

130-097-658

This kit isolated PMN directly from murine bone marrow and gave great results (easier and faster than FACS, excellent results).

Experimental Design and Results Summary

Application

Isolation of PMN - negative selection

Starting Material

Bone marrow (murine)

Protocol Overview

Followed protocol

Tips

Did all incubations at 4C (cold room or fridge, not on ice).

Results Summary

Easy to use, routinely got over 90% purity. There were a lot of neutrophils left in the column, so the yield could have been higher, but I am happy with the purity. I added extra washes to get more of the neutrophils out, but I think some loss of neutrophils is inevitable (unless you have very few cells / pure culture you put into the column).

Additional Notes

None

Image Gallery

Summary

The Good

Easy and effective, untouched neutrophil isolation. Faster than FACS sorting

The Bad

Need MACS column and magnet? I personally like this better than FACS though.

The Bottom Line

In the image, I have three samples, one is bone marrow (preisolation), one is the PMN enrichment (elution from column), and the one called "depleted" is the cells that were left in the column, which includes neutophils. Overall, very happy with the kit, as well as customer service from MACS.

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