biotinylated goat anti human lap polyclonal antibody Search Results


mouse cd45 antibody  (Bio-Techne corporation)
93
Bio-Techne corporation mouse cd45 antibody
Mouse Cd45 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse cd45 antibody/product/Bio-Techne corporation
Average 93 stars, based on 1 article reviews
mouse cd45 antibody - by Bioz Stars, 2026-02
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biotinylated goat anti-human fasl polyclonal antibodies  (PeproTech)
90
PeproTech biotinylated goat anti-human fasl polyclonal antibodies
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Biotinylated Goat Anti Human Fasl Polyclonal Antibodies, supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/biotinylated goat anti-human fasl polyclonal antibodies/product/PeproTech
Average 90 stars, based on 1 article reviews
biotinylated goat anti-human fasl polyclonal antibodies - by Bioz Stars, 2026-02
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biotinylated goat anti-human apociii polyclonal antibodies k74170b  (Biodesign International Inc)
90
Biodesign International Inc biotinylated goat anti-human apociii polyclonal antibodies k74170b
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Biotinylated Goat Anti Human Apociii Polyclonal Antibodies K74170b, supplied by Biodesign International Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/biotinylated goat anti-human apociii polyclonal antibodies k74170b/product/Biodesign International Inc
Average 90 stars, based on 1 article reviews
biotinylated goat anti-human apociii polyclonal antibodies k74170b - by Bioz Stars, 2026-02
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biotinylated goat anti-human apo(aii) polyclonal antibody  (Biodesign International Inc)
90
Biodesign International Inc biotinylated goat anti-human apo(aii) polyclonal antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Biotinylated Goat Anti Human Apo(Aii) Polyclonal Antibody, supplied by Biodesign International Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/biotinylated goat anti-human apo(aii) polyclonal antibody/product/Biodesign International Inc
Average 90 stars, based on 1 article reviews
biotinylated goat anti-human apo(aii) polyclonal antibody - by Bioz Stars, 2026-02
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human trem2 biotinylated antibody  (Bio-Techne corporation)
99
Bio-Techne corporation human trem2 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Trem2 Biotinylated Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human trem2 biotinylated antibody/product/Bio-Techne corporation
Average 99 stars, based on 1 article reviews
human trem2 biotinylated antibody - by Bioz Stars, 2026-02
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human/mouse cxcl12/sdf-1 biotinylated antibody  (Bio-Techne corporation)
94
Bio-Techne corporation human/mouse cxcl12/sdf-1 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human/Mouse Cxcl12/Sdf 1 Biotinylated Antibody, supplied by Bio-Techne corporation, 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/result/human/mouse cxcl12/sdf-1 biotinylated antibody/product/Bio-Techne corporation
Average 94 stars, based on 1 article reviews
human/mouse cxcl12/sdf-1 biotinylated antibody - by Bioz Stars, 2026-02
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human epcam/trop-1 biotinylated antibody  (Bio-Techne corporation)
99
Bio-Techne corporation human epcam/trop-1 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Epcam/Trop 1 Biotinylated Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human epcam/trop-1 biotinylated antibody/product/Bio-Techne corporation
Average 99 stars, based on 1 article reviews
human epcam/trop-1 biotinylated antibody - by Bioz Stars, 2026-02
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human gata-4 biotinylated antibody  (Bio-Techne corporation)
93
Bio-Techne corporation human gata-4 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Gata 4 Biotinylated Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human gata-4 biotinylated antibody/product/Bio-Techne corporation
Average 93 stars, based on 1 article reviews
human gata-4 biotinylated antibody - by Bioz Stars, 2026-02
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human il-8/cxcl8 biotinylated antibody  (Bio-Techne corporation)
94
Bio-Techne corporation human il-8/cxcl8 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Il 8/Cxcl8 Biotinylated Antibody, supplied by Bio-Techne corporation, 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/result/human il-8/cxcl8 biotinylated antibody/product/Bio-Techne corporation
Average 94 stars, based on 1 article reviews
human il-8/cxcl8 biotinylated antibody - by Bioz Stars, 2026-02
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biotinylated polyclonal goat anti-human taci antibody 500-p166gbt  (PeproTech)
90
PeproTech biotinylated polyclonal goat anti-human taci antibody 500-p166gbt
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Biotinylated Polyclonal Goat Anti Human Taci Antibody 500 P166gbt, supplied by PeproTech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/biotinylated polyclonal goat anti-human taci antibody 500-p166gbt/product/PeproTech
Average 90 stars, based on 1 article reviews
biotinylated polyclonal goat anti-human taci antibody 500-p166gbt - by Bioz Stars, 2026-02
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human ccl17/tarc biotinylated antibody  (Bio-Techne corporation)
90
Bio-Techne corporation human ccl17/tarc biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Ccl17/Tarc Biotinylated Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human ccl17/tarc biotinylated antibody/product/Bio-Techne corporation
Average 90 stars, based on 1 article reviews
human ccl17/tarc biotinylated antibody - by Bioz Stars, 2026-02
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human igf-i/igf-1 biotinylated antibody  (Bio-Techne corporation)
90
Bio-Techne corporation human igf-i/igf-1 biotinylated antibody
(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with <t>biotinylated</t> WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a <t>polyclonal</t> anti-human <t>FasL</t> antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.
Human Igf I/Igf 1 Biotinylated Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human igf-i/igf-1 biotinylated antibody/product/Bio-Techne corporation
Average 90 stars, based on 1 article reviews
human igf-i/igf-1 biotinylated antibody - by Bioz Stars, 2026-02
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(A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with biotinylated WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a polyclonal anti-human FasL antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.

Journal: PLoS ONE

Article Title: The bioactivity of soluble Fas ligand is modulated by key amino acids of its stalk region

doi: 10.1371/journal.pone.0253260

Figure Lengend Snippet: (A) Competition assay. Serial 3-fold dilutions of either WT or 8-site mutated sFasL were mixed with biotinylated WT sFasL (70 ng/ml) and incubated for 2 hours in 96-well plates pre-coated with recombinant human soluble Fas (250 ng/ml). Biotin was detected using HRP conjugated streptavidin; (B) Fas binding assay. Serial 3-fold dilutions of wild type (WT) or mut-sFasL were incubated for 2 hours in wells coated with recombinant human soluble Fas, 5 μg/ml. The sFasL bound to Fas was detected using a polyclonal anti-human FasL antibody. The mut sFasL binding curve was shifted to the left. n = 3. (C) Jurkat cells were incubated in 96-well plates at a concentration of 1 x 10 4 cells/well. Different molar ratios of the Fas-activating antibody CH-11 and mut-sFasL were added, and caspase 3/7 activity measured after 5 hours. The mut-sFasL did not inhibit the activity of CH11. Data represent the results from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Sidak’s post hoc analysis; (A), WT-sFasL compared to mut-sFasL at the same concentrations; (B), comparisons made to no competitor condition for each molecule; (C) Comparisons made to unmixed antibody (Ab). Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.

Article Snippet: For the Fas-FasL binding assay, we used recombinant human soluble Fas receptors (PeproTech, Rocky Hill, NJ) and biotinylated goat anti-human FasL polyclonal antibodies (PeproTech Cat# 500-P184bt-50ug, RRID:AB_148184).

Techniques: Competitive Binding Assay, Incubation, Recombinant, Binding Assay, Concentration Assay, Activity Assay

Serial 3-fold dilutions of FLAG tagged mut-sFasL were mixed with biotinylated WT sFasL (60 ng). The mixtures were incubated at room temperature for 15 minutes and applied to streptavidin-coated wells. FLAG tags were detected with HRP anti-FLAG mAb. There was a dose-dependent increase in FLAG signal in wells incubated with the biotinylated WT sFasL and the mut-sFasL-FLAG mixture, indicating formation of complexes of WT sFasL and mut-sFasL. The mut-sFasL-FLAG without biotin did not bind to the streptavidin-coated plates. Data shown as ratio of the absorbance at 450 nm of each experimental condition to that of media only. Data were generated from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Dunnet’s post hoc analysis; comparisons were made to 0 ng/mL for each condition. Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.

Journal: PLoS ONE

Article Title: The bioactivity of soluble Fas ligand is modulated by key amino acids of its stalk region

doi: 10.1371/journal.pone.0253260

Figure Lengend Snippet: Serial 3-fold dilutions of FLAG tagged mut-sFasL were mixed with biotinylated WT sFasL (60 ng). The mixtures were incubated at room temperature for 15 minutes and applied to streptavidin-coated wells. FLAG tags were detected with HRP anti-FLAG mAb. There was a dose-dependent increase in FLAG signal in wells incubated with the biotinylated WT sFasL and the mut-sFasL-FLAG mixture, indicating formation of complexes of WT sFasL and mut-sFasL. The mut-sFasL-FLAG without biotin did not bind to the streptavidin-coated plates. Data shown as ratio of the absorbance at 450 nm of each experimental condition to that of media only. Data were generated from three separate experiments, each done in duplicate, and were analyzed by 2-way ANOVA with Dunnet’s post hoc analysis; comparisons were made to 0 ng/mL for each condition. Each dot represents individual data, lines represent means ± SD. * = P < 0.05; ** = P<0.01, ***P<0.001.

Article Snippet: For the Fas-FasL binding assay, we used recombinant human soluble Fas receptors (PeproTech, Rocky Hill, NJ) and biotinylated goat anti-human FasL polyclonal antibodies (PeproTech Cat# 500-P184bt-50ug, RRID:AB_148184).

Techniques: Incubation, Generated