streptavidin coated biacore biosensor chips Search Results


t200 instrument  (Cytiva Europe)
96
Cytiva Europe t200 instrument
T200 Instrument, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Biacore+T200/pm27133466-271-11-13
Average 96 stars, based on 1 article reviews
t200 instrument - by Bioz Stars, 2026-09
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streptavidin sensor chip  (Biacore)
86
Biacore streptavidin sensor chip
Streptavidin Sensor Chip, supplied by Biacore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/chips+s+sa+series/birkholz_alysia_marie__2015__immune_and_structural_studies_of_synthetic_invariant_natural_killer_t_cell_glycosphingolipid-923-9-12
Average 86 stars, based on 1 article reviews
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sensor streptavidin chips  (Valiant Co Ltd)
94
Valiant Co Ltd sensor streptavidin chips
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Sensor Streptavidin Chips, supplied by Valiant Co Ltd, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Streptavidin/pmc05123071-170-17-10
Average 94 stars, based on 1 article reviews
sensor streptavidin chips - by Bioz Stars, 2026-09
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series s sensor chip sa biosensor  (Cytiva Europe)
95
Cytiva Europe series s sensor chip sa biosensor
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Series S Sensor Chip Sa Biosensor, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Series+S+Sensor+Chip+SA%2C+pack+of+1/pmc06974358-424-13-19
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series s sensor chip sa biosensor - by Bioz Stars, 2026-09
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streptavidin coated sensor chips  (Biacore)
86
Biacore streptavidin coated sensor chips
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Streptavidin Coated Sensor Chips, supplied by Biacore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/biacore+chip+sensor/10__1021_slash_ja211628j-60-16-19
Average 86 stars, based on 1 article reviews
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biacore sa chips  (Biacore)
86
Biacore biacore sa chips
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Biacore Sa Chips, supplied by Biacore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/biacore+surface/10__1042_slash_bj20090839-112-0-0
Average 86 stars, based on 1 article reviews
biacore sa chips - by Bioz Stars, 2026-09
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streptavidin derivatised gold chip  (Biacore)
86
Biacore streptavidin derivatised gold chip
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Streptavidin Derivatised Gold Chip, supplied by Biacore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/chip+cm3+s+sensor+series/pm42168605-164-25-31
Average 86 stars, based on 1 article reviews
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streptavidin conjugated biacore sensor chips  (Cytiva Europe)
95
Cytiva Europe streptavidin conjugated biacore sensor chips
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Streptavidin Conjugated Biacore Sensor Chips, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Sensor+Chip+SA/pmc11414566-44-50-57
Average 95 stars, based on 1 article reviews
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sensor chip sa  (Cytiva Europe)
95
Cytiva Europe sensor chip sa
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Sensor Chip Sa, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Sensor+Chip+SA%2C+pack+of+3/pmc06172828-292-8-11
Average 95 stars, based on 1 article reviews
sensor chip sa - by Bioz Stars, 2026-09
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streptavidin coated sensor chip  (Biacore)
86
Biacore streptavidin coated sensor chip
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Streptavidin Coated Sensor Chip, supplied by Biacore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/antibodycell+killing/pm21193695-48-30-34
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streptavidin coated sensor chip - by Bioz Stars, 2026-09
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streptavidin biosensor chip  (Cytiva Europe)
95
Cytiva Europe streptavidin biosensor chip
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Streptavidin Biosensor Chip, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/streptavidin+coated+biacore+biosensor+chips/Series+S+Sensor+Chip+SA%2C+pack+of+3/pmc04171027-402-9-12
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cm5 sensor chip  (Thermo Fisher)
99
Thermo Fisher cm5 sensor chip
( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different <t>streptavidin</t> chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.
Cm5 Sensor Chip, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different streptavidin chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.

Journal: Nature Communications

Article Title: The Apaf-1 apoptosome induces formation of caspase-9 homo- and heterodimers with distinct activities

doi: 10.1038/ncomms13565

Figure Lengend Snippet: ( a ) Apaf-1 (300 nM) was incubated with C9-p35/12, C9-p35/p12-CIVSM, ProC9-TM, ProC9-TM-CIVSM or ProC9-TM-F404D (200 nM)±Cc/dATP (2 mM each) for 30 min at 37 °C in a final volume of 200 μl. Following separation by Superose-6 gel filtration chromatography, protein fractions corresponding to monomeric proteins (M), apoptosome complexes (Ap) and protein aggregates (Ag) were pooled, precipitated with trichloroacetic acid, separated by SDS–PAGE and immunoblotted for Apaf-1 and C9. The experiments were repeated three times with similar results. ( b , c ) Binding affinities of Apaf-1 CARD for C9 prodomain, ProC9-TM, ProC9-TM-F404D, C9-p35/p12, C9-p35/p12-CIVSM and C9/8 were determined by SPR, as described in the Methods section. Analysis of all proteins was repeated five or more times using at least two different streptavidin chips, and each value represents the mean±s.e.m. Left panel: raw curves at 2 μM for each protein; right panel: sensograms for each of the full-length C9 proteins were normalized to their own peak heights to highlight differences in k off . ( d ) Apoptosome complexes were reconstituted with ProC9-TM or C9-p35/p12 (200 nM)±catalytically inactive ProC9* or ProC9*-F404D (67.5 nM–2 μM) and then assayed for LEHDase activity. Each protein was assayed at least four times, and each value represents the mean±s.e.m. ( e ) Wild-type mouse embryonic fibroblasts (MEFs) expressing either ProC9* or ProC9*-F404D were exposed to heat shock for 1 h or ultraviolet irradiation for 4 min and assayed for cell death at 24 h by propidium iodide (PI) staining/flow cytometry . The experiment was repeated four times, and each bar represents the mean±s.e.m. * P <0.05, analysis of variance, Student–Newman–Keuls post hoc analysis.

Article Snippet: LEHD.amc (catalogue# 03AMC154) and DEVD.amc (catalogue# 03AMC138) were acquired from MP Biomedicals, and L -DOPA and Biacore sensor streptavidin chips (catalogue# BR-1000-32) were obtained from Acros and GE Biosciences, respectively.

Techniques: Incubation, Filtration, Chromatography, SDS Page, Binding Assay, Activity Assay, Expressing, Irradiation, Staining, Flow Cytometry