Efficient Myelin Removal from Adult Mouse Brain for Subsequent Astrocyte Isolation

SUNY UB
Research Scientist

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Company:

Miltenyi Biotec

Product Name:

Myelin Isolation Beads, human, mouse, rat

Catalog Number:

130-104-257

We use Myelin Isolation Beads during the preparation of single-cell suspensions from adult mouse brain tissue for subsequent immunomagnetic isolation of astrocytes. In our workflow, myelin depletion is performed before ACSA-2–based magnetic separation, and the resulting astrocyte-enriched fraction is used for downstream molecular analyses, including Western blotting.

Experimental Design and Results Summary

Application

Myelin removal from dissociated adult mouse brain tissue prior to immunomagnetic isolation of ACSA-2–positive astrocytes.

Starting Material

Dissociated prefrontal cortex tissue from adult mice.

Tips

Check the expiration date immediately upon delivery, particularly when ordering the 4 mL bottle. Only a relatively small amount of reagent is needed per isolation, so laboratories performing adult brain isolations intermittently should consider whether they will be able to use the bottle before it expires.

Results Summary

The Myelin Isolation Beads efficiently removed myelin from dissociated adult mouse brain preparations and produced a cleaner cell suspension for subsequent magnetic separation. We used the myelin-depleted fraction for ACSA-2–based astrocyte isolation and obtained astrocyte-enriched samples suitable for downstream protein analysis. Western blot validation of the isolated fraction showed enrichment of the astrocytic marker S100β with minimal neuronal and microglial contamination, supporting the effectiveness of the overall isolation workflow.

DOI or PMID #

10.1126/sciadv.adt2527

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Summary

The Good

Efficient myelin removal from adult mouse brain preparations, straightforward integration into the Miltenyi magnetic separation workflow, and good compatibility with subsequent ACSA-2–based astrocyte isolation.

The Bad

The major drawback is the short remaining shelf life upon delivery. We received the 4 mL bottle with only about two months remaining before the expiration date. Because only a small amount is needed for each isolation, it can be difficult to use the entire bottle before it expires, resulting in considerable reagent waste.

The Bottom Line

Myelin Isolation Beads are a useful and effective reagent for preparing adult mouse brain cell suspensions for subsequent immunomagnetic cell isolation. They integrate well into our ACSA-2 astrocyte isolation workflow and help produce clean preparations suitable for downstream molecular analyses. The main limitation in our experience was the very short remaining shelf life of the 4 mL bottle upon delivery, which should be considered when planning experiments and choosing how much reagent to order.

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