Cell surface biotinylation

Brenda H. Welter; Heather A. Walters; Lesly A. Temesvari

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Cell surface biotinylation

BW Brenda H. Welter

HW Heather A. Walters

LT Lesly A. Temesvari

This method is extracted from research article: PLoS One, Mar 2020

Reduced expression of a rhomboid protease, EhROM1, correlates with changes in the submembrane distribution and size of the Gal/GalNAc lectin subunits in the human protozoan parasite, Entamoeba histolytica

DOI: 10.1371/journal.pone.0219870

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Control and T-EhROM1-s cells (6.0 × 10⁵) were surface biotinylated and purified by avidin affinity chromatography using the Pierce Cell Surface Protein Isolation Kit (Pierce Biotechnology, Rockford, IL, USA) according to manufacturer’s specifications. Flow-through fractions and biotinylated surface proteins, captured by avidin affinity chromatography, were resolved by SDS-PAGE and analyzed for Hgl, Lgl, Igl and actin by western blotting and densitometry as described above.

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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