Native BAD-1 Binding to Heparin-agarose

Materials and Reagents

1. Heparin-agarose resin (Sigma-Aldrich, catalog number: H6508 ) (prior to use it is washed 3x in five volumes of tricine buffer to eliminate free heparin)
   
2. 10 μg of BAD-1 [purified according to the method of Brandhorst et al. (2005)]
   
3. 25 microliters of soluble medical-grade sodium heparin for injection (50 mg/ml) (Elkins-Sinn Inc)
   
4. Tricine buffer (Sigma-Aldrich, catalog number: T0377 ) (see Recipes)
   

Equipment

1. Accuspin micro17 microcentrifuge (Thermo Fisher Scientific)
   
2. Nanodrop ND1000 spectrophotometer
   

Procedure