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HOOK™ Cell Surface Protein Isolation

Catalog Description Size Price(USD) Qty
786-316
HOOK™ Cell Surface Protein Isolation 5 Reactions
$258.00

HOOK™ Cell Surface Protein Isolation

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. 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.
 
Following 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.
 

HOOK™ sulfo-NHS-SS-Biotin   

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: 7-9

 

Following 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.

 


Features

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

 


Applications

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

 

 

Protocol
786-316
Material Safety Data Sheet
786-316
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Technical Literature
Mass Spectrometry Sample Prep Handbook A guide to the preparation of protein samples for Mass Spectrometry, including protein extraction, clean-up and peptide generation.
Protein Electrophoresis Handbook A guide to 1D and 2D protein electrophoresis products, including protein markers, electrophoresis buffers, 2D electrophoresis reagents, clean-up reagents and stains. The guide also offers protein sample preparation products.
Protein Labeling & Conjugation
Protein Purification Handbook
Sample Preparation Handbook For Lysis Buffers, Fractionation, Dialysis, Protein Concentration and Enrichment
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