HOOK™ Cell Surface Protein Isolation, 5 Reactions, 786-316, G-BIOSCIENCES (CAT#: STEM-GT-3026-LGZ)

The HOOK™ Cell Surface Protein Isolation Kit uses G-Biosciences HOOK™ Biotin Labeling and Purification Technology in combination with our Mammalian Cell PE LB™ Lysis Buffer to conveniently label cell surface proteins and isolate them for further analysis, including Western blotting.

Mammalian cells, adherent or non-adherent, are first labeled with HOOK™ Sulfane-NHS-SS-Biotin. HOOK™ Sulfo-NHS-SS-Biotin is a water-soluble amine-reactive biotinylation reagent with N-hydroxysulfosuccinimide (Sulfo-NHS) ester (see table). The addition of a charged sulfonate (SO3-) to the N-hydroxysuccinimide ring of the sulfo-NHS ester renders it soluble in water but prevents it from permeating the plasma membrane. Solubility and impermeability to the plasma membrane make HOOK™ Sulfo-NHS-SS-Biotin ideal for studying cell surface proteins as it only reacts with protein molecules on the outer surface of the plasma membrane. Another advantage of HOOK™ Sulfane-NHS-SS-Biotin is the presence of a disulfide bond in the spacer arm. The disulfide bond allows the biotin moiety to be cleaved from the protein, making its interaction with avidin/streptavidin reversible.

After labeling, cells are lysed with Mammalian PE LB™, a buffered lysate that uses mild non-ionic detergents to enhance protein extraction and stability, and the cell lysate is applied to a Streptavidin agarose column. Unlabeled intracellular proteins are washed away and biotinylated cell surface proteins are released by reduction of the disulfide bond with DTT.

The kit provides all necessary reagents and buffers for convenience and increased reproducibility. The kit is compatible with a variety of mammalian cells and can be used to compare differences between treated and untreated cells as well as between different cell lines. The kit provides enough reagents for 5 experiments, each consisting of 4 90-95% confluent T-755cm2 flasks.

Size: 5 Reactions
Hook™ sulfo-NHS-SS-Biotin
Molecular weight: 606.69
Spacer (Å): 24.3
Reactive group: sulfo-naphthalene sulfonate
Membrane permeability: no
Water Solubility: Yes
Divisible/Reversible: Yes
Reaction pH value: 7-9

Complete cell surface biotin labeling and isolation.
Suitable for a wide selection of mammalian cells.

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