Identification of Specific Relaxin: MATERIALS AND METHODS(2)
Each tissue sample was cut into slices (0.5-3 cm3) and individually placed in peel-A-way plastic embedding molds (Polysciences, Inc., Warrington, PA). The tissues were frozen with Tissue-Tek O.C.T. embedding compound (Miles Scientific, Elkhart, IN) in liquid nitrogen and stored at -70°C until sectioning. Frozen sections (8 ^m) were cut on an HR Mark II cryostat (Slee Medical Equipment Ltd., London, England) at – 20°C and thaw-mounted on microscope slides coated with 0.2% poly-L-lysine (Mr 300 000). Tissue slides were incubated in anhydrous acetone at -30°C for 3 h. buy ventolin inhalers
Preparation and Characterization of Biotinylated Relaxin
Porcine relaxin was isolated as described by Sherwood and O’Byrne and biotinylated by a modification of the procedure described by Bullesbach and Schwabe. In brief, porcine relaxin was dissolved in 0.2 M N-methylmorpholine-HCl buffer (pH 7.5) at a final concentration of 2 ^mol/ml. To supply the biotinylating reagent in excess, 10 molar equivalents of biotinyl-e-aminocaproic acid-N-hydroxy succinimide ester (Sigma, St. Louis, MO) in dimethylformamide at a concentration of 100 ^mol/ml were added to the relaxin.
Tags: Cells, Female, Human, Relaxin