HOOK™ Cell Surface Protein Isolatio...

HOOK™ Cell Surface Protein Isolation

Product Information

Product Overview:

HOOK™ Cell Surface Protein Isolation kit uses G-Biosciences HOOK™ biotin labeling and purification technology in conjunction 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™ Sulfo-NHS-SS-Biotin (see figure 1). HOOK™ Sulfo-NHS-SS-Biotin is a water-soluble, amine reactive biotinylation reagent that has a N-hydroxysulfosuccinimide (sulfo-NHS) ester (see table). The addition of a charged sulfonate (SO3-) on the N-hydroxysuccinimide ring of the sulfo-NHS esters results in its solubility in water, but prevents it permeating plasma membranes. The solubility and impermeability to plasma membranes makes HOOK™ Sulfo-NHS-SS-Biotin ideal for studying cell surface proteins as it will only react with the protein molecules on the outer surface of plasma membranes. An additional advantage of HOOK™ Sulfo-NHS-SS-Biotin is the presence of a disulfide bond in the spacer arm. The disulfide bond permits the cleavage of the biotin moiety from the protein, making its interaction with avidin/ streptavidin reversible. HOOK™ sulfo-NHS-SS-Biotin (figure 2) MOLECULAR WEIGHT 606.69 SPACER ARM (Å) 24.3 REACTIVE GROUP sulfo-NHS-ester MEMBRANE PERMEABLE NO WATER SOLUBLE YES CLEAVABLE/ REVERSIBLE YES REACTIVE pH 7-9Following labeling, cells are lysed with Mammalian Cell PE LB™, a buffered lysis solution that employs a mild non-ionic detergent for enhanced extraction and stability of proteins, and the cell lysate is applied to a Streptavidin agarose column. Unlabeled intracellular proteins are washed away and the biotin labeled cell surface proteins are then released by reduction of the disulfide bond with DTT. The kit is supplied with all the necessary reagents and buffers for convenience and improved reproducibility. The kit is compatible with a wide variety of mammalian cells and can be used to compare treated and untreated cells and differences between different cell lines. This kit is supplied with sufficient reagents for five experiments, with each experiment consisting of four 90-95% confluent T-75cm2 flasks.

Application:

For the isolation of cell surface proteins. For the study of cell surface trafficking. For the study of receptor:ligand interactions.

Features:

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

Catalog#: 786-316
LID#: 4323P
Quantity :
Order by Phone or Fax: 1.800.316.3081

Specifications

Warehouse Storage Temp4C
DescriptionFor the biotin labeling and subsequent isolation of cell surface proteins
Unit Size5 Reactions
Safety StatementsFor Research Use Only
Sample AvailabilityYes
SynonymHOOK Cell Surface Protein Isolation, biotin, sulfo-NHS-SS-Biotin, streptavidin, purification

Shipping & Handling

Estimated Ship Time: 2 Business Days

Shipped In: Ambient

Hazardous Material: No

Storage

Shelf Life: 12 MONTHS

Storage Temperature: 4C

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