Sulfosuccinimidyl-6- hexanoate,

Active ester stock solution A dissolvelO mg of sulfosuccinimidyl-6-(biotini-mide)hexanoate (BIO-NHS, Pierce) in 1 ml of DMF. 

Sulfo-SANPAH Sulfosuccinimidyl-6-(4 -azido-2 -nitrophenylamino)hexanoate MW 492.39 18.2 A... 

One hundred milligrams of sulfosuccinimidyl-6- (biotinimide) hexanoate (NHS-LC-biotin) (Pierce, Rockford, IL) is dissolved in 40 ml of 50 mM borate buffer (pH 8.0). To this solution is added 30 mg of tyramine hydrochloride (Sigma Chemical Company, St. Louis, MO). The solution is stirred overnight at room temperature and filtered (0.45 p,m filter). The final biotinylated tyramine concentration is 7 xM. Before application, the solution is diluted 1 160 in Tris-HCl buffer (pH 7.6) containing 0.03% H202. 

Amankwa et al. (1992) (129) successfully derivatized 3-aminopropyl-triethoxysilane (2%) onto the inner wall of fused silica capillary. Following this treatment, the capillary was perfused with a 5.0 mg/mL solution of NHS-LC-biotin (sulfosuccinimidyl 6-(biotinamideo)hexanoate), and it was subsequently treated with avidin in 0.05 M sodium phosphate buffer (pH 7.4). Finally, the avidin-coated capillary was rinsed with distilled water and then treated with a 10 mg/mL solution of biotin-labeled trypsin in sodium phosphate buffer by gravity flow (shovm in scheme 8.7). 

A water-soluble version of this cross-linker also exists. Sulfosuccinimidyl-6-(4 -azido-2 -nitrophenylamino)hexanoate (sulfo-SANPAH) contains the negatively charged sulfonate group on its NHS ring, lending greater hydrophilicity to the compound. 

Sulfosuccinimidyl 6-(biotinamido) hexanoate, also called Sulfo-NHS-LG-biotin (Pierce and Warriner (UK) Ltd.)... 

Add 100 mg of sulfosuccinimidyl 6-(biotinamido) hexanoate and 30 mg of tyramine-HCl to 40 ml of borate buffer, then stir gently overnight. 

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