Thermo Scientific EZ-Link Desthiobiotinylation and Sedimentation Kits are a complete set of kits for biotinylation of purified proteins Reagents for purifying interacting complexes using streptavidin agarose resin under mild elution conditions.

EZ-Link Desthiobiotinylation and Sedimentation Kit Features:

•Sulfo-NHS-LC-Desthiobiotin – Labeled with biotin-like reagents proteins, this reagent binds specifically to streptavidin but also elutes easily under mild conditions
• Sedimentation Assay – The protocol and reagents are simple and optimized for Label proteins and recover (purify) intact functional protein interaction complexes
• Complete kit – includes labeling reagents, desalting columns for preparing bait proteins, and strands for performing sedimentation assays Mycin Avidin Sepharose and Elution Buffer

This all-in-one labeling and purification kit uses Sulfo-NHS-LC-desthiobiotin, an amine-reactive NHS ester reagent , which efficiently labels purified or mixed protein samples for use as bait proteins in capture and purified protein interactions. The desthiobiotin tag of the labeled protein is conjugated to streptavidin, which is highly specific but easily elutes under very mild conditions (i.e., in a manner that is competitively displaced by conventional free biotin). This system efficiently and gently captures and isolates (sediments) specific protein-interacting complexes from cell lysates using streptavidin agarose beads. This complete kit includes ready-to-use 1mg vials of desthiobiotinylation reagent, a desalting column for processing labeled proteins, and high-capacity streptavidin agarose for easily performing multiple microcentrifuge-scale sedimentation assays. and buffer. Identify novel interactions between known proteins (bait proteins) and previously undiscovered target proteins (prey proteins), or confirm interactions between known bait proteins and prey proteins.

Desthiobiotin is a monocyclic, sulfur-free biotin analog that is more specific for binding to streptavidin than biotin. The sexes are almost equal, but the affinity is lower (1/Kd = 10¹¹ vs. 10¹⁵M respectively). Therefore, desthiobiotinylated bait proteins and their interacting partners can be easily and specifically washed from streptomycin affinity resins using mild conditions based on competitive displacement with free biotin. Come out. For sedimentation assays using biological samples, this soft release characteristic of desthiobiotin also helps to minimize co-purification of endogenous biotinylated molecules when elution of target protein complexes with free biotin Still bound to strepavidin. The modified avidin-biotin affinity system also eliminates the need for harsh elution conditions that may separate the complex and/or damage the target protein or cells. Desthiobiotin technology is ideal for working with native or recombinant proteins that do not express fusion tags or for isolating capture proteins under native conditions, such as when targeting intact cells or cell surface proteins.

Sulfo-NHS-LC-desthiobiotin is a biotin variant that can be activated to a sulfo-NHS ester with a long chain (LC) spacer arm that is covalently linked to the desthiobiotin group Primary amine (-NH2) that labels proteins or other molecules. No-Weigh format (1 mg resealable vials) eliminates the difficulty of weighing small amounts of reagents and protects unused reagents from hydrolysis as much as possible.

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