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Red blood cells were centrifuged and washed in isotonic buffered salt solution, collected in ice cold lysis buffer (20 mM Hepes, pH 7.4, 200 μM CaCl2, 0.25 M sucrose, and proteinase inhibitors (Roche)), homogenized by a loosely fitting dounce homogenizer and additional passages through a 22 gauge needle. A 50 % iodixanol solution was established by mixing Optiprep solution (60 % iodixanol; Sigma) with buffer (120 mM Hepes, pH 7.4, 1.2 mM CaCl2, 0.25 M sucrose). Seven parts of this 50 % iodixanol solution were mixed with thirteen parts of the red blood cell solution, generating a gradient solution containing 17.5 % iodixanol. The mixture was filled into a centrifuge tube underlayed by cushions of 30 % and 35 % iodixanol and centrifuged for 3 hours at 270,000 × g in a SW41 Ti swing out rotor (Beckman) to achieve a nearly linear density gradient (approximately 1.06 g/ml to 1.20 g/ml) [24]. Fractions of 0.6 ml were collected from the top of the gradient by a 1 ml syringe without using a needle and immediately frozen in liquid nitrogen for further use."}
craft-ca-core-dev
{"project":"craft-ca-core-dev","denotations":[{"id":"T7207","span":{"begin":13,"end":22},"obj":"CHEBI:31705"},{"id":"T7208","span":{"begin":44,"end":49},"obj":"NCBITaxon:9606"},{"id":"T7209","span":{"begin":50,"end":65},"obj":"CL:0000232"},{"id":"T7210","span":{"begin":54,"end":59},"obj":"UBERON:0000178"},{"id":"T7211","span":{"begin":98,"end":108},"obj":"PR:000004080"},{"id":"T7212","span":{"begin":142,"end":157},"obj":"CL:0000232"},{"id":"T7213","span":{"begin":146,"end":151},"obj":"UBERON:0000178"},{"id":"T7214","span":{"begin":244,"end":249},"obj":"GO:0019835"},{"id":"T7215","span":{"begin":264,"end":269},"obj":"CHEBI:46756"},{"id":"T7216","span":{"begin":286,"end":291},"obj":"CHEBI:3312"},{"id":"T7217","span":{"begin":300,"end":307},"obj":"CHEBI:17992"},{"id":"T7218","span":{"begin":455,"end":464},"obj":"CHEBI:31705"},{"id":"T7219","span":{"begin":465,"end":473},"obj":"CHEBI:75958"},{"id":"T7220","span":{"begin":509,"end":517},"obj":"CHEBI:75958"},{"id":"T7221","span":{"begin":524,"end":533},"obj":"CHEBI:31705"},{"id":"T7222","span":{"begin":562,"end":567},"obj":"CHEBI:46756"},{"id":"T7223","span":{"begin":584,"end":589},"obj":"CHEBI:3312"},{"id":"T7224","span":{"begin":598,"end":605},"obj":"CHEBI:17992"},{"id":"T7225","span":{"begin":633,"end":642},"obj":"CHEBI:31705"},{"id":"T7226","span":{"begin":643,"end":651},"obj":"CHEBI:75958"},{"id":"T7227","span":{"begin":690,"end":704},"obj":"CL:0000232"},{"id":"T7228","span":{"begin":694,"end":699},"obj":"UBERON:0000178"},{"id":"T7229","span":{"begin":705,"end":713},"obj":"CHEBI:75958"},{"id":"T7230","span":{"begin":737,"end":745},"obj":"CHEBI:75958"},{"id":"T7231","span":{"begin":764,"end":773},"obj":"CHEBI:31705"},{"id":"T7232","span":{"begin":779,"end":786},"obj":"CHEBI:60004"},{"id":"T7233","span":{"begin":861,"end":870},"obj":"CHEBI:31705"}],"text":"Self forming iodixanol density gradients of human red blood cells\nThe subcellular distribution of annexin A7 was addressed according to [24]. Red blood cells were centrifuged and washed in isotonic buffered salt solution, collected in ice cold lysis buffer (20 mM Hepes, pH 7.4, 200 μM CaCl2, 0.25 M sucrose, and proteinase inhibitors (Roche)), homogenized by a loosely fitting dounce homogenizer and additional passages through a 22 gauge needle. A 50 % iodixanol solution was established by mixing Optiprep solution (60 % iodixanol; Sigma) with buffer (120 mM Hepes, pH 7.4, 1.2 mM CaCl2, 0.25 M sucrose). Seven parts of this 50 % iodixanol solution were mixed with thirteen parts of the red blood cell solution, generating a gradient solution containing 17.5 % iodixanol. The mixture was filled into a centrifuge tube underlayed by cushions of 30 % and 35 % iodixanol and centrifuged for 3 hours at 270,000 × g in a SW41 Ti swing out rotor (Beckman) to achieve a nearly linear density gradient (approximately 1.06 g/ml to 1.20 g/ml) [24]. Fractions of 0.6 ml were collected from the top of the gradient by a 1 ml syringe without using a needle and immediately frozen in liquid nitrogen for further use."}
2_test
{"project":"2_test","denotations":[{"id":"12925238-7978280-8102424","span":{"begin":137,"end":139},"obj":"7978280"},{"id":"12925238-7978280-8102425","span":{"begin":1037,"end":1039},"obj":"7978280"}],"text":"Self forming iodixanol density gradients of human red blood cells\nThe subcellular distribution of annexin A7 was addressed according to [24]. Red blood cells were centrifuged and washed in isotonic buffered salt solution, collected in ice cold lysis buffer (20 mM Hepes, pH 7.4, 200 μM CaCl2, 0.25 M sucrose, and proteinase inhibitors (Roche)), homogenized by a loosely fitting dounce homogenizer and additional passages through a 22 gauge needle. A 50 % iodixanol solution was established by mixing Optiprep solution (60 % iodixanol; Sigma) with buffer (120 mM Hepes, pH 7.4, 1.2 mM CaCl2, 0.25 M sucrose). Seven parts of this 50 % iodixanol solution were mixed with thirteen parts of the red blood cell solution, generating a gradient solution containing 17.5 % iodixanol. The mixture was filled into a centrifuge tube underlayed by cushions of 30 % and 35 % iodixanol and centrifuged for 3 hours at 270,000 × g in a SW41 Ti swing out rotor (Beckman) to achieve a nearly linear density gradient (approximately 1.06 g/ml to 1.20 g/ml) [24]. Fractions of 0.6 ml were collected from the top of the gradient by a 1 ml syringe without using a needle and immediately frozen in liquid nitrogen for further use."}
craft-ca-core-ex-dev
{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T7234","span":{"begin":13,"end":22},"obj":"CHEBI:31705"},{"id":"T7235","span":{"begin":44,"end":49},"obj":"NCBITaxon:9606"},{"id":"T7236","span":{"begin":50,"end":65},"obj":"CL:0000232"},{"id":"T7237","span":{"begin":54,"end":59},"obj":"UBERON:0000178"},{"id":"T7238","span":{"begin":60,"end":65},"obj":"CL_GO_EXT:cell"},{"id":"T7239","span":{"begin":70,"end":81},"obj":"GO_UBERON_EXT:cellular_component_or_cell_part"},{"id":"T7240","span":{"begin":73,"end":81},"obj":"CL_GO_EXT:cell"},{"id":"T7241","span":{"begin":98,"end":108},"obj":"PR_EXT:000004080"},{"id":"T7242","span":{"begin":142,"end":157},"obj":"CL:0000232"},{"id":"T7243","span":{"begin":146,"end":151},"obj":"UBERON:0000178"},{"id":"T7244","span":{"begin":152,"end":157},"obj":"CL_GO_EXT:cell"},{"id":"T7245","span":{"begin":198,"end":206},"obj":"CHEBI_CHMO_EXT:buffer_process"},{"id":"T7246","span":{"begin":207,"end":220},"obj":"CHEBI_EXT:saline_solution"},{"id":"T7247","span":{"begin":244,"end":249},"obj":"GO:0019835"},{"id":"T7248","span":{"begin":250,"end":256},"obj":"CHEBI_CHMO_EXT:buffer_solution"},{"id":"T7249","span":{"begin":264,"end":269},"obj":"CHEBI:46756"},{"id":"T7250","span":{"begin":286,"end":291},"obj":"CHEBI:3312"},{"id":"T7251","span":{"begin":300,"end":307},"obj":"CHEBI:17992"},{"id":"T7252","span":{"begin":313,"end":334},"obj":"CHEBI_GO_EXT:peptidase_or_protease_or_proteinase_inhibitor"},{"id":"T7253","span":{"begin":412,"end":420},"obj":"GO_EXT:biological_movement_or_translocation_process"},{"id":"T7254","span":{"begin":455,"end":464},"obj":"CHEBI:31705"},{"id":"T7255","span":{"begin":465,"end":473},"obj":"CHEBI:75958"},{"id":"T7256","span":{"begin":509,"end":517},"obj":"CHEBI:75958"},{"id":"T7257","span":{"begin":524,"end":533},"obj":"CHEBI:31705"},{"id":"T7258","span":{"begin":547,"end":553},"obj":"CHEBI_CHMO_EXT:buffer_solution"},{"id":"T7259","span":{"begin":562,"end":567},"obj":"CHEBI:46756"},{"id":"T7260","span":{"begin":584,"end":589},"obj":"CHEBI:3312"},{"id":"T7261","span":{"begin":598,"end":605},"obj":"CHEBI:17992"},{"id":"T7262","span":{"begin":633,"end":642},"obj":"CHEBI:31705"},{"id":"T7263","span":{"begin":643,"end":651},"obj":"CHEBI:75958"},{"id":"T7264","span":{"begin":690,"end":704},"obj":"CL:0000232"},{"id":"T7265","span":{"begin":694,"end":699},"obj":"UBERON:0000178"},{"id":"T7266","span":{"begin":700,"end":704},"obj":"CL_GO_EXT:cell"},{"id":"T7267","span":{"begin":705,"end":713},"obj":"CHEBI:75958"},{"id":"T7268","span":{"begin":737,"end":745},"obj":"CHEBI:75958"},{"id":"T7269","span":{"begin":764,"end":773},"obj":"CHEBI:31705"},{"id":"T7270","span":{"begin":779,"end":786},"obj":"CHEBI:60004"},{"id":"T7271","span":{"begin":861,"end":870},"obj":"CHEBI:31705"},{"id":"T7272","span":{"begin":1180,"end":1188},"obj":"CHEBI_EXT:nitrogen"}],"text":"Self forming iodixanol density gradients of human red blood cells\nThe subcellular distribution of annexin A7 was addressed according to [24]. Red blood cells were centrifuged and washed in isotonic buffered salt solution, collected in ice cold lysis buffer (20 mM Hepes, pH 7.4, 200 μM CaCl2, 0.25 M sucrose, and proteinase inhibitors (Roche)), homogenized by a loosely fitting dounce homogenizer and additional passages through a 22 gauge needle. A 50 % iodixanol solution was established by mixing Optiprep solution (60 % iodixanol; Sigma) with buffer (120 mM Hepes, pH 7.4, 1.2 mM CaCl2, 0.25 M sucrose). Seven parts of this 50 % iodixanol solution were mixed with thirteen parts of the red blood cell solution, generating a gradient solution containing 17.5 % iodixanol. The mixture was filled into a centrifuge tube underlayed by cushions of 30 % and 35 % iodixanol and centrifuged for 3 hours at 270,000 × g in a SW41 Ti swing out rotor (Beckman) to achieve a nearly linear density gradient (approximately 1.06 g/ml to 1.20 g/ml) [24]. Fractions of 0.6 ml were collected from the top of the gradient by a 1 ml syringe without using a needle and immediately frozen in liquid nitrogen for further use."}