Thermo Scientific™ Pierce™ Biotinylated Protein Interaction Pull-Down Kit

The kit makes it possible to perform an efficient pull-down assay with high protein recovery and reproducibility. This protocol uses immobilized streptavidin to capture a biotinylated protein and a spin column format that enables complete gel retention during each capture, wash and elution step. When the biotinylated bait protein is immobilized, it can withstand stringent washes and remains bound during prey protein elution, which results in an isolated prey protein without bait protein contamination. This technique is ideal when using native or recombinant proteins that are not expressed with a fusion tag. Additionally, prey protein is isolated from the immobilized bait protein using native binding conditions and nondenaturing elution conditions. The Biotinylated Protein Interaction Pull-Down Kit is designed to teach the method to first-time users and to increase ease-of-use and convenience for experienced researchers.

Highlights:

• Biotin pull-down – purifies protein interactors of any biotinylated protein or biotin-tagged ligand

• Complete kit – provides all components and detailed protocol for purifying protein:protein interactions

• No special equipment needed – uses common laboratory equipment and reagents (e.g., microcentrifuge)

• Spin column efficiency – microcentrifuge spin columns facilitate microcentrifuge spin columns facilitate simple and efficient manipulation of streptavidin agarose beads, including simple processing of multiple samples

Includes:

Streptavidin agarose resin, TBS pack, biotin blocking buffer, wash buffer, elution buffer, spin columns and collection tubes

Recommended for:

Discover a new protein interaction complex using a biotinylated protein as the bait to capture a putative binding partner from a cell lysate or other sample; Confirm expression of a known protein interaction target in a cell lysate; Determine the ability of two purified proteins to interact (bind) in various buffer conditions; Extract protein:protein interaction information from in vitro transcription/ translation lysates