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The immunoblotting was carried out as described previously ( 11; 14 ). Briefly, the cells were lysed by adding 20 μl of protein A and 3 mm EDTA containing 50 dithiothreitol, 20 mm triton with 10 EDTA, and 5 mm EDTA. The samples were centrifuged at 1,750 g for 10 min and the supernatant left for 10 min. Then the cell Citalopram 20mg $218.33 - $0.61 Per pill pellet was subjected to 2 ml of 10% sucrose and centrifuged at 10,000 rpm for 5 min. After the supernatant was discarded, pellet subjected to 1 ml of 10% triton with 10 mm EDTA. The supernatant was again discarded and the pellet was centrifuged for 5 min at 10,000 rpm to separate the membrane from cells. membranes were then rinsed with 0.1% Triton X-100, centrifuged, and resolved on 14.5% polyacrylamide gels.
The total intracellular calcium generic drugstore hiring was measured by using the CaCl 2 -sensor [Fisher Scientific ECL-450, Milwaukee, WI; calibrated according to the manufacturer's specifications ( 10 )]. Briefly, 20,000 μl of phosphate buffer, pH 7.4, 5.6, and 200 μl of 0.3% bovine serum albumin in 10 μl of sodium phosphate buffer saline containing 10 mm sodium EDTA and nmol/l glucose was incubated at 37°C for 1 h. The order citalopram online uk absorbance was read at 540 nm. The intracellular calcium concentration measured was in mmol/L.
Plasma corticosterone concentrations and hormone levels were measured by using commercially available kits (Wako Chemicals GmbH, Germany). Briefly, the corticosterone concentration, as determined by using a commercially licensed kit, was measured using a Biosan-Vaceral C.
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