PMC:3585731 / 54479-55698 JSON TXT

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testone

{"project":"testone","denotations":[{"id":"T27054","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27053","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

pmc-enju-pas

bionlp-st-ge-2016-test-proteins

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T27078","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27077","span":{"begin":168,"end":172},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T27320","span":{"begin":327,"end":331},"obj":"http://www.uniprot.org/uniprot/P01375"},{"id":"T27319","span":{"begin":168,"end":172},"obj":"http://www.uniprot.org/uniprot/P01375"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

UBERON-AE

{"project":"UBERON-AE","denotations":[{"id":"T27062","span":{"begin":802,"end":806},"obj":"http://purl.obolibrary.org/obo/UBERON_0001913"},{"id":"T27061","span":{"begin":820,"end":825},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"},{"id":"T27060","span":{"begin":229,"end":234},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"},{"id":"T27059","span":{"begin":195,"end":200},"obj":"http://purl.obolibrary.org/obo/UBERON_0001977"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

GO-BP

{"project":"GO-BP","denotations":[{"id":"T27080","span":{"begin":1088,"end":1095},"obj":"http://purl.obolibrary.org/obo/GO_0023052"},{"id":"T27079","span":{"begin":278,"end":284},"obj":"http://purl.obolibrary.org/obo/GO_0040007"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

GO-MF

{"project":"GO-MF","denotations":[{"id":"T27083","span":{"begin":1208,"end":1218},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T27082","span":{"begin":963,"end":973},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T27081","span":{"begin":895,"end":905},"obj":"http://purl.obolibrary.org/obo/GO_0003823"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T27096","span":{"begin":1208,"end":1218},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T27095","span":{"begin":963,"end":973},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T27094","span":{"begin":895,"end":905},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T27093","span":{"begin":1208,"end":1218},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T27092","span":{"begin":963,"end":973},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T27091","span":{"begin":895,"end":905},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T27090","span":{"begin":1112,"end":1121},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T27089","span":{"begin":778,"end":786},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T27088","span":{"begin":470,"end":474},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T27087","span":{"begin":218,"end":223},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T27086","span":{"begin":108,"end":113},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T27085","span":{"begin":53,"end":58},"obj":"http://purl.obolibrary.org/obo/GO_0005623"},{"id":"T27084","span":{"begin":33,"end":38},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

sentences

{"project":"sentences","denotations":[{"id":"T27074","span":{"begin":1097,"end":1219},"obj":"Sentence"},{"id":"T27073","span":{"begin":1029,"end":1096},"obj":"Sentence"},{"id":"T27072","span":{"begin":887,"end":1028},"obj":"Sentence"},{"id":"T27071","span":{"begin":730,"end":886},"obj":"Sentence"},{"id":"T27070","span":{"begin":609,"end":729},"obj":"Sentence"},{"id":"T27069","span":{"begin":524,"end":608},"obj":"Sentence"},{"id":"T27068","span":{"begin":446,"end":523},"obj":"Sentence"},{"id":"T27067","span":{"begin":333,"end":445},"obj":"Sentence"},{"id":"T27066","span":{"begin":174,"end":332},"obj":"Sentence"},{"id":"T27065","span":{"begin":92,"end":173},"obj":"Sentence"},{"id":"T27064","span":{"begin":0,"end":91},"obj":"Sentence"},{"id":"T342","span":{"begin":0,"end":91},"obj":"Sentence"},{"id":"T343","span":{"begin":92,"end":173},"obj":"Sentence"},{"id":"T344","span":{"begin":174,"end":332},"obj":"Sentence"},{"id":"T345","span":{"begin":333,"end":445},"obj":"Sentence"},{"id":"T346","span":{"begin":446,"end":523},"obj":"Sentence"},{"id":"T347","span":{"begin":524,"end":608},"obj":"Sentence"},{"id":"T348","span":{"begin":609,"end":665},"obj":"Sentence"},{"id":"T349","span":{"begin":666,"end":729},"obj":"Sentence"},{"id":"T350","span":{"begin":730,"end":886},"obj":"Sentence"},{"id":"T351","span":{"begin":887,"end":952},"obj":"Sentence"},{"id":"T352","span":{"begin":953,"end":1028},"obj":"Sentence"},{"id":"T353","span":{"begin":1029,"end":1096},"obj":"Sentence"},{"id":"T354","span":{"begin":1097,"end":1219},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

simple1

{"project":"simple1","denotations":[{"id":"T27100","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27099","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

BioNLP16_DUT

{"project":"BioNLP16_DUT","denotations":[{"id":"T27318","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27317","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

BioNLP16_Messiy

{"project":"BioNLP16_Messiy","denotations":[{"id":"T27326","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27325","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

DLUT931

{"project":"DLUT931","denotations":[{"id":"T27322","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27321","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

bionlp-st-ge-2016-test-ihmc

{"project":"bionlp-st-ge-2016-test-ihmc","denotations":[{"id":"T27345","span":{"begin":40,"end":58},"obj":"Entity"},{"id":"T27344","span":{"begin":524,"end":546},"obj":"Entity"},{"id":"T27343","span":{"begin":887,"end":905},"obj":"Entity"},{"id":"T27342","span":{"begin":135,"end":149},"obj":"Entity"},{"id":"T27341","span":{"begin":773,"end":777},"obj":"Entity"},{"id":"T27340","span":{"begin":810,"end":839},"obj":"Protein"},{"id":"T27339","span":{"begin":623,"end":634},"obj":"Protein"},{"id":"T27366","span":{"begin":333,"end":363},"obj":"Binding"},{"id":"T27365","span":{"begin":0,"end":90},"obj":"Binding"},{"id":"T27364","span":{"begin":582,"end":607},"obj":"Entity"},{"id":"T27363","span":{"begin":218,"end":223},"obj":"Entity"},{"id":"T27362","span":{"begin":1050,"end":1062},"obj":"Entity"},{"id":"T27361","span":{"begin":773,"end":786},"obj":"Entity"},{"id":"T27360","span":{"begin":953,"end":973},"obj":"Entity"},{"id":"T27359","span":{"begin":18,"end":59},"obj":"Entity"},{"id":"T27358","span":{"begin":108,"end":113},"obj":"Entity"},{"id":"T27357","span":{"begin":1170,"end":1172},"obj":"Entity"},{"id":"T27356","span":{"begin":333,"end":338},"obj":"Entity"},{"id":"T27355","span":{"begin":1204,"end":1218},"obj":"Entity"},{"id":"T27354","span":{"begin":372,"end":376},"obj":"Entity"},{"id":"T27353","span":{"begin":300,"end":308},"obj":"Entity"},{"id":"T27352","span":{"begin":470,"end":482},"obj":"Entity"},{"id":"T27351","span":{"begin":1050,"end":1053},"obj":"Protein"},{"id":"T27350","span":{"begin":1112,"end":1121},"obj":"Entity"},{"id":"T27349","span":{"begin":153,"end":172},"obj":"Protein"},{"id":"T27348","span":{"begin":294,"end":331},"obj":"Protein"},{"id":"T27347","span":{"begin":739,"end":752},"obj":"Entity"},{"id":"T27346","span":{"begin":835,"end":838},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

bionlp-st-ge-2016-spacy-parsed

bionlp-st-ge-2016-test-tees

{"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T27332","span":{"begin":835,"end":838},"obj":"Protein"},{"id":"T27331","span":{"begin":820,"end":833},"obj":"Protein"},{"id":"T27330","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27329","span":{"begin":297,"end":308},"obj":"Protein"},{"id":"T27328","span":{"begin":168,"end":172},"obj":"Protein"},{"id":"T27327","span":{"begin":138,"end":149},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

test3

{"project":"test3","denotations":[{"id":"T27058","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27057","span":{"begin":168,"end":172},"obj":"Protein"},{"id":"T27056","span":{"begin":327,"end":331},"obj":"Protein"},{"id":"T27055","span":{"begin":168,"end":172},"obj":"Protein"}],"text":"For Western blot, 4×105 adherent cells (1×106 Jurkat cells) were seeded into 35 mm2 dishes. After 24–48 hr, cells were stimulated with 30 µM zVAD.fmk or 10 ng/ml mouse TNFα. For treatments under serum free conditions, cells were serum starved for 24 hr prior to the addition of growth factors, 20 µM zVAD.fmk or 10 ng/ml mouse TNFα. Cells were harvested in 1×RIPA buffer (Cell Signaling) supplemented with 50 µg/ml phenylmethanesulfonylfluoride. After brief sonication, cell lysates were spun down for 15 min at 14,000×rpm. Protein concentrations were measured using the Pierce 660 nm Assay Reagent (Pierce). Equal amounts of proteins were boiled for 5 min at 95°C. Western blotting was performed according to standard protocols. Briefly, SDS-PAGE gels were transferred to PVDF membrane, blocked in 3% milk or 5% bovine serum albumin (BSA) in TBST buffer for 30 min at room temperature. Primary antibodies were incubated in 5%BSA/TBST overnight at 4°C. Secondary antibodies were incubated in TBST for 30 min at room temperature. Luminata (Millipore) ECL reagents were used to develop the signals. In some cases, membranes were stripped using OneMinute stripping buffer (GM Biosciences) and reprobed with new antibodies."}

PMC:3585731 / 54479-55698 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:3585731 / 54479-55698 JSON TXT