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Serum was diluted 1 1 with double concentrated TBST.
After coating period, saliva was aspirated from each well and replaced with 500 μL of BHI broth (double concentrated) and 500 μL of saliva.
Protein digestion, peptide isolation and desalination were carried out according to Toyofuku et al. [ 30] with double concentrated NH4HCO3 buffer, 30 min of carbamidomethylation and only one step of formic acid extraction (5% (v/v) formic acid).
Briefly, 1 mg collagen I was dissolved in 1 mL of ice cold 10 mM acetic acid and was mixed with an equal volume of 1 mg (s GAG derivative/mL dissolved in ice cold double concentrated fibrillogenesis buffer (50 mM sodium dihydrogenphosphate and 11 mM potassium dihydrogenphosphate, pH 7.4).
After separation, proteins were transferred to a PVDF membrane (Immun-BlotTM, Bio-Rad CA, USA) using an electrophoresis tank filled with a double concentrated running buffer without SDS supplemented with 10%% (v/v) methanol for 45 min at 100 V constant voltage.
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Popular aims were raking (firing a broadside the length of an enemy ship from across the bow or stern) or doubling (concentrating force by putting ships on both sides of the enemy line).
(Serves 4) 1 tbsp sunflower oil, 3p 1 onion, chopped, 11p 6 free-range sausages, £2.79 400g tin baked beans, 22p 400g tin chopped tomatoes, 31p 142g double concentrate tomato puree, 35p 220ml bitter, 13p 1 tsp winter herbs (rosemary, oregano or thyme, or mixed herbes de Provence), 3p 1 stock cube, 2p Heat the oil in a large saute pan and add the onion.
The pellet was then extracted with 15 μl of double-concentrate SDS-PAGE sample loading buffer (pH 6.8).
Before, Clijsters had to quit doubles to concentrate on singles.
Though both have since stopped playing doubles to concentrate on singles, they have found each other across the net from one another less frequently.
The polar vortex, typically, is a single-cell or double-cell region concentrated at polar latitudes.
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