Works to Cleave Alpha 2-3 Sialylations, but Is Not Very Efficient

Princeton
Molecular Biology
Graduate Student

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Company:

New England Biolabs

Product Name:

α2-3,6,8 Neuraminidase

Catalog Number:

P0720S

This enzyme was used to assess whether the SLeX moiety could be degraded to prevent selectin interaction with intact cancer cells. The enzyme worked moderately well over a two hour digestion, but results were messy.

Experimental Design and Results Summary

Application

Gkycobiology/Flow cytometry

Starting Material

Fut5 modified cancer cells

Protocol Overview

Cells were lifted from plates and digested with enzyme at 1:1000 for 2 hours at 37°C. They were then probed with recombinant Selectin reagents to assess the extent of SLex degradation by interaction measured via flow cytometry.

Tips

This may work better with free-floating proteins in a specialized solution, doesn't work great in PBS

Results Summary

Results showed that while the enzyme could cleave the sialic acid, it did not do so entirely and resulted in a messy signal.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

It is the only enzyme that performs this reaction

The Bad

Not a high turnover enzyme or may have other biophysical limitations

The Bottom Line

Works moderately, but not great for sensitive life science applications.

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