PMC:3320587 / 13599-14748 JSON TXT

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pmc-enju-pas

bionlp-st-ge-2016-test-proteins

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T6004","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T6003","span":{"begin":506,"end":509},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

bionlp-st-ge-2016-uniprot

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T6235","span":{"begin":688,"end":693},"obj":"http://www.uniprot.org/uniprot/O94916"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

GO-BP

{"project":"GO-BP","denotations":[{"id":"T6005","span":{"begin":53,"end":58},"obj":"http://purl.obolibrary.org/obo/GO_0019835"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

GO-MF

{"project":"GO-MF","denotations":[{"id":"T6007","span":{"begin":781,"end":789},"obj":"http://purl.obolibrary.org/obo/GO_0003823"},{"id":"T6006","span":{"begin":694,"end":702},"obj":"http://purl.obolibrary.org/obo/GO_0003823"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

GO-CC

{"project":"GO-CC","denotations":[{"id":"T6013","span":{"begin":781,"end":789},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T6012","span":{"begin":694,"end":702},"obj":"http://purl.obolibrary.org/obo/GO_0042571"},{"id":"T6011","span":{"begin":781,"end":789},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T6010","span":{"begin":694,"end":702},"obj":"http://purl.obolibrary.org/obo/GO_0019815"},{"id":"T6009","span":{"begin":425,"end":433},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T6008","span":{"begin":19,"end":23},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

sentences

{"project":"sentences","denotations":[{"id":"T6000","span":{"begin":1032,"end":1149},"obj":"Sentence"},{"id":"T5999","span":{"begin":867,"end":1031},"obj":"Sentence"},{"id":"T5998","span":{"begin":617,"end":866},"obj":"Sentence"},{"id":"T5997","span":{"begin":444,"end":616},"obj":"Sentence"},{"id":"T5996","span":{"begin":361,"end":443},"obj":"Sentence"},{"id":"T5995","span":{"begin":231,"end":360},"obj":"Sentence"},{"id":"T5994","span":{"begin":13,"end":230},"obj":"Sentence"},{"id":"T5993","span":{"begin":0,"end":12},"obj":"Sentence"},{"id":"T83","span":{"begin":0,"end":12},"obj":"Sentence"},{"id":"T84","span":{"begin":13,"end":230},"obj":"Sentence"},{"id":"T85","span":{"begin":231,"end":360},"obj":"Sentence"},{"id":"T86","span":{"begin":361,"end":443},"obj":"Sentence"},{"id":"T87","span":{"begin":444,"end":616},"obj":"Sentence"},{"id":"T88","span":{"begin":617,"end":866},"obj":"Sentence"},{"id":"T89","span":{"begin":867,"end":1031},"obj":"Sentence"},{"id":"T90","span":{"begin":1032,"end":1149},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

simple1

{"project":"simple1","denotations":[{"id":"T6015","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T6014","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

BioNLP16_DUT

{"project":"BioNLP16_DUT","denotations":[{"id":"T6489","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T6488","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

BioNLP16_Messiy

{"project":"BioNLP16_Messiy","denotations":[{"id":"T6243","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T6242","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

DLUT931

{"project":"DLUT931","denotations":[{"id":"T6241","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T6240","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

bionlp-st-ge-2016-test-ihmc

{"project":"bionlp-st-ge-2016-test-ihmc","denotations":[{"id":"T6520","span":{"begin":1093,"end":1148},"obj":"Entity"},{"id":"T6519","span":{"begin":489,"end":499},"obj":"Entity"},{"id":"T6518","span":{"begin":759,"end":789},"obj":"Protein"},{"id":"T6517","span":{"begin":842,"end":865},"obj":"Entity"},{"id":"T6516","span":{"begin":231,"end":239},"obj":"Entity"},{"id":"T6515","span":{"begin":673,"end":710},"obj":"Entity"},{"id":"T6514","span":{"begin":934,"end":974},"obj":"Protein"},{"id":"T6513","span":{"begin":96,"end":104},"obj":"Entity"},{"id":"T6512","span":{"begin":77,"end":105},"obj":"Entity"},{"id":"T6511","span":{"begin":388,"end":442},"obj":"Entity"},{"id":"T6510","span":{"begin":361,"end":368},"obj":"Entity"},{"id":"T6509","span":{"begin":164,"end":173},"obj":"Entity"},{"id":"T6508","span":{"begin":712,"end":717},"obj":"Protein"},{"id":"T6507","span":{"begin":568,"end":578},"obj":"Entity"},{"id":"T6506","span":{"begin":96,"end":104},"obj":"Entity"},{"id":"T6505","span":{"begin":324,"end":332},"obj":"Protein"},{"id":"T6504","span":{"begin":114,"end":123},"obj":"Entity"},{"id":"T6503","span":{"begin":583,"end":594},"obj":"Entity"},{"id":"T6502","span":{"begin":298,"end":319},"obj":"Entity"},{"id":"T6501","span":{"begin":474,"end":477},"obj":"Entity"},{"id":"T6500","span":{"begin":155,"end":182},"obj":"Protein"},{"id":"T6499","span":{"begin":503,"end":517},"obj":"Protein"},{"id":"T6498","span":{"begin":800,"end":805},"obj":"Protein"},{"id":"T6497","span":{"begin":101,"end":104},"obj":"Entity"},{"id":"T6496","span":{"begin":511,"end":516},"obj":"Protein"},{"id":"T6495","span":{"begin":125,"end":137},"obj":"Entity"},{"id":"T6494","span":{"begin":24,"end":32},"obj":"Entity"},{"id":"T6493","span":{"begin":183,"end":192},"obj":"Entity"},{"id":"T6492","span":{"begin":388,"end":442},"obj":"Entity"},{"id":"T6491","span":{"begin":606,"end":609},"obj":"Protein"},{"id":"T6490","span":{"begin":405,"end":410},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

bionlp-st-ge-2016-spacy-parsed

bionlp-st-ge-2016-test-tees

{"project":"bionlp-st-ge-2016-test-tees","denotations":[{"id":"T6245","span":{"begin":688,"end":693},"obj":"Protein"},{"id":"T6244","span":{"begin":174,"end":192},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

testone

{"project":"testone","denotations":[{"id":"T5988","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T5987","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

test3

{"project":"test3","denotations":[{"id":"T5992","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T5991","span":{"begin":506,"end":509},"obj":"Protein"},{"id":"T5990","span":{"begin":829,"end":832},"obj":"Protein"},{"id":"T5989","span":{"begin":506,"end":509},"obj":"Protein"}],"text":"Western blot\nWhole cell extracts were collected with lysis buffer containing 150 mM NaCl, 50 mM Tris–HCl, pH 7.5, 1% Triton, 10% glycerol, and 1 tablet of Complete EDTA-free Protease Inhibitor Cocktail (Roche) per 25 ml of buffer. Extracts were boiled for 5 min in 1× Laemmli sample buffer with 5% v/v 2-mercaptoethanol and proteins were separated by SDS-PAGE. The gel was transferred to a nitrocellulose Trans-Blot Transfer Membrane (BioRad). The blot was then blocked for 1 h at 37°C in a solution of 4% BSA (Sigma) and 0.1% Tween-20 (BioRad) in a buffer containing 50 mM Tris and 150 mM NaCl at pH 7.6 (BSA/TBST). Primary incubation was carried out with a1∶200 dilution of rabbit anti-NFAT5 antibody (H-300) (Santa Cruz Biotechnology) and a 1∶500 dilution of goat anti-Lamin-B1 antibody (sc-6217; Santa Cruz Biotechnology) in BSA/TBST for 2 h at room temperature. The blot was washed 3×5 min in TBST and incubated in 1∶6000 donkey anti-goat-HRP (Santa Cruz Biotechnology) or goat anti-rabbit-HRP (BioRad) as appropriate for 1 h. The blot was again washed 3×5 min in TBST and developed with SuperSignal West Pico Chemiluminescent Reagent (Pierce)."}

PMC:3320587 / 13599-14748 JSONTXT

Annnotations TAB JSON ListView MergeView

PMC:3320587 / 13599-14748 JSON TXT