Thermo Scientific™ Pierce™ Cell Surface Protein Isolation Kit

Description

Thermo Scientific™ Cell Surface Protein Isolation Kit is a complete set of reagents for selective biotinylation and subsequent purification of mammalian cell surface proteins to the exclusion of intracellular proteins.

The kit efficiently labels proteins that have accessible lysine residues and sufficient extracellular exposure.The isolation procedure uses a cell-impermeable, cleavable biotinylation reagent (Sulfo-NHS-SS-Biotin) to label exposed primary amines of proteins on the surface of intact adherent or suspension cells. Treated cells are then harvested and lysed, and the labeled surface proteins are affinity-purified using Thermo Scientific NeutrAvidin Agarose Resin. The isolated cell surface proteins contain a small, nonreactive tag of the originally labeled primary amines but are no longer biotinylated (biotin remains bound to the resin). The kit contains sufficient reagents for eight experiments, each involving four T75 flasks of confluent cells.

Highlights:

• Isolates cell surface proteins – reduces complexity of total cellular protein

• Efficiently recovers labeled proteins – cleavable biotin allows for nearly 100% recovery of isolated cell surface proteins

• Convenience – all reagents are provided in one kit, along with complete instructions for labeling, cell lysis and purification of cell surface membrane proteins

• Western blotting applications – proteins recovered in SDS-PAGE buffer are loaded directly onto polyacrylamide gels

• Robust system – protocol designed for diverse cell lines, including NIH 3T3, HeLa, C6 and A431

Includes:

Biotinylation reagent, cell lysis reagent, agarose affinity resin, various buffers and microcentrifuge spin columns