Cell Surface Protein Isolation Kit provides a simple and efficient method for the isolation of cell surface proteins. In this method, cells are first labeled with Sulfo-NHS-SS-Biotin, an amine-reactive, thiol-cleavable, biotinylation reagent. Cells are subsequently lysed and the labeled cell surface proteins are isolated using Streptavidin beads. The bound proteins are then released from beads by incubating with DTT solution. Biotinylation reagent is cell-membrane-impermeable with an extended spacer arm to reduce steric hindrances associated with streptavidin binding. This convenient kit provides all the required components for optimal labeling and isolation of the cell surface proteins.
Cell surface proteins play a major role in signal transduction, cell adhesion, transport and serve as diagnostic and pharmacological targets.
Isolated cell surface proteins can be used for various downstream applications such as Western Blotting or other structural and functional studies.
Store kit at 4°C. Briefly spin small vials prior to opening. Read entire protocol before performing the assay.PBS Tablet: Dissolve one PBS Tablet in 100 ml of ddH2O to make 1X PBS solution. Sterile filter the solution and store at 4°C.TBS Tablet: Dissolve in 100 ml of ddH2O to make 1X TBS solution. Sterile filter the solution and store at 4°C.Sulfo-NHS-SS-Biotin: Each vial is good for two reactions. Reconstitute each vial with 2 ml of 1XPBS. Pipette up and down to completely dissolve the powder. Make working solution of Sulfo-NHS-SS-Biotin by adding 2 ml reconstituted solution to 18 ml of ice cold 1XPBS. Notes: a. Always reconstitute Sulfo-NHS-SS-Biotin vial just before use. b. Optional: Sulfo-NHS-SS-Biotin can also be reconstituted in anhydrous DMSO or DMF. Reconstitute the powder in 100 μl of DMSO or DMF and add to 19.9 ml of ice cold 1XPBS.DTT: Aliquot and store at -20°C. Use within 2 months.
Quenching Solution: 10 mlLysis Buffer: 6.5 mlWash Buffer: 12 mlStreptavidin Beads: 1.5 mlPBS Tablet: 3TBS Tablet: 1Sulfo-NHS-SS-Biotin: 5 VialsDTT: 100 µl
Compatible Sample Types:
Adherent and suspension cells
Features & Benefits:
• Simple and efficient method for the isolation of cell surface proteins• Includes all required components for optimal labeling and isolation of the cell surface proteins