Thermo Scientific™ alkaline phosphatase-conjugated NeutrAvidin Protein is a specially-prepared form of avidin that decreases background in biotin-binding.
Features include: • Near-neutral isoelectric point—pI = 6.3, more neutral than native avidin • Nearly devoid of glycosylation—decreased possibility of lectin binding compared to native avidin • No RYD recognition sequence—no known off-target binding domains like streptavidin • Affordable—significantly less expensive than streptavidin
NeutrAvidin Protein is deglycosylated native avidin from egg whites. Removal of the excess carbohydrate by an exclusive process yields a protein with a more neutral isoelectric point and less nonspecific binding properties. Conjugated NeutrAvidin Protein provides exceptional performance in western blot, ELISA, and IHC applications that require biotin-binding probes. Assay specificity, sensitivity, and signal-to-noise ratios with NeutrAvidin Protein are generally equivalent or better than with the significantly more expensive streptavidin.
Avidin is a glycoprotein found in the egg white and tissues of birds, reptiles, and amphibia. The biotin-binding protein contains four identical subunits having a combined mass of 67,000-68,000 daltons. Removing the glycosylation from avidin yields NeutrAvidin Protein with a mass of 60,000 daltons. Carbohydrate-based lectin binding is reduced to undetectable levels, yet biotin-binding affinity is retained. NeutrAvidin Protein offers the advantages of a neutral pI to minimize nonspecific adsorption, along with lysine residues that remain available for derivatization or other customized conjugation. NeutrAvidin Protein yields the lowest nonspecific binding among the known biotin-binding proteins. The specific activity for biotin binding is approximately 14 µg/mg of protein, which is near the theoretical maximum activity.
Applications: • In situ hybridization—NeutrAvidin-AP Conjugate is currently being used by the Allen Institute for Brain Science to create the Allen Brain Atlas. • Western blotting—use NeutrAvidin Protein conjugates (HRP or AP) instead of streptavidin conjugates for detection of biotinylated target proteins or biotinylated primary or secondary antibodies in western blots. • ELISA—NeutrAvidin Protein conjugates (HRP or AP) replace corresponding streptavidin conjugates for direct or indirect sandwich ELISA detection schemes involving biotin-labeled proteins and antibodies. Depending on the specific assay components (e.g., blocking buffers, etc.), NeutrAvidin Protein conjugates provide similar or better assay performance measured as signal-to-noise ratio.