phenylmethylsulfonyl fluoride Search Results


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  • 99
    Millipore phenylmethylsulfonyl fluoride
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethylsulfonyl Fluoride, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 24760 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Beyotime phenylmethanesulfonyl fluoride
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethanesulfonyl Fluoride, supplied by Beyotime, used in various techniques. Bioz Stars score: 93/100, based on 2072 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Beyotime phenylmethanesulfonyl fluoride pmsf
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethanesulfonyl Fluoride Pmsf, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 651 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher phenylmethylsulfonyl fluoride
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethylsulfonyl Fluoride, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 2011 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Boehringer Mannheim phenylmethylsulfonyl fluoride
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethylsulfonyl Fluoride, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 713 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore m phenylmethylsulfonyl fluoride
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    M Phenylmethylsulfonyl Fluoride, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1608 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher phenylmethylsulfonyl fluoride pmsf
    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by <t>phenylmethylsulfonyl</t> fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P
    Phenylmethylsulfonyl Fluoride Pmsf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 961 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad phenylmethylsulfonyl fluoride
    Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM <t>phenylmethylsulfonyl</t> fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.
    Phenylmethylsulfonyl Fluoride, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 435 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc phenylmethylsulfonyl fluoride
    Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM <t>phenylmethylsulfonyl</t> fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.
    Phenylmethylsulfonyl Fluoride, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 414 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Beyotime protease inhibitor phenylmethanesulfonyl fluoride
    Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM <t>phenylmethylsulfonyl</t> fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.
    Protease Inhibitor Phenylmethanesulfonyl Fluoride, supplied by Beyotime, used in various techniques. Bioz Stars score: 91/100, based on 475 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology phenylmethylsulfonyl fluoride
    Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM <t>phenylmethylsulfonyl</t> fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.
    Phenylmethylsulfonyl Fluoride, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Amresco phenylmethanesulfonyl fluoride
    Effect of CdTe QDs of different sizes on autophagy in yeast at different time points. Notes: Cells transformed with a plasmid encoding GFP-Atg8 were treated with ( A ) 50 nmol/L CdTe QDs or 100 ng/mL rapamycin or ( B ) <t>phenylmethanesulfonyl</t> fluoride (PMSF), which was added at 0 hour and 16 hours to reach a final concentration of 2 mmol/L, and photographed at 4 hours, 8 hours, 16 hours, and 42 hours using fluorescence microscopy. Scale bars: 5 µm. Abbreviations: CdTe QDs, cadmium telluride quantum dots; G-CdTe, green-emitting CdTe; O-CdTe, orange-emitting CdTe.
    Phenylmethanesulfonyl Fluoride, supplied by Amresco, used in various techniques. Bioz Stars score: 92/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Nacalai phenylmethylsulfonyl fluoride
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethylsulfonyl Fluoride, supplied by Nacalai, used in various techniques. Bioz Stars score: 92/100, based on 205 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Beijing Solarbio Science phenylmethylsulfonyl fluoride
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethylsulfonyl Fluoride, supplied by Beijing Solarbio Science, used in various techniques. Bioz Stars score: 90/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Branson Ultrasonics phenylmethanesulfonyl fluoride
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethanesulfonyl Fluoride, supplied by Branson Ultrasonics, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fisher Scientific phenylmethylsulfonyl fluoride
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethylsulfonyl Fluoride, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 93/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Valiant phenylmethanesulfonyl fluoride
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethanesulfonyl Fluoride, supplied by Valiant, used in various techniques. Bioz Stars score: 92/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc phenylmethanesulfonyl fluoride pmsf
    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF <t>phenylmethylsulfonyl</t> fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown
    Phenylmethanesulfonyl Fluoride Pmsf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P

    Journal: Redox Biology

    Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction

    doi: 10.1016/j.redox.2018.07.005

    Figure Lengend Snippet: MMP inhibitors prevent the increases in vascular gelatinolytic activity, in ROS concentrations, and in lucigenin chemiluminescence after intraluminal incubation of rabbit aorta with MMP-2. (A) Representative photomicrographs of aortic in situ zymography experiments showing that MMP-2 gelatinolytic activity was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The arteries were incubated with the inhibitors together with MMP-2 (16 nM) intraluminally for 30 min. (B) Quantification of aortic gelatinolytic activity measured in experiments described in A. (C) Representative photomicrographs of aortic sections incubated with DHE. Red fluorescence reflects ROS production and shows that incubation with MMP-2 (16 nM) intraluminally for 30 min increased ROS production, and this effect was totally inhibited by phenanthroline (Phe, 100 µM) or doxycycline (Doxy, 100 µM), but not by phenylmethylsulfonyl fluoride (PMSF, 100 µM). The experiments were carried out as described in A. (D), Quantification of aortic fluorescence of DHE measured in experiments described in C. (E), Quantification of lucigenin chemiluminescence measured in fresh aortas after intraluminal incubation with MMP-2 (16 nM) or vehicle for 30 min in absence or presence of MMP inhibitors, doxycycline (Doxy 100 µM) or GM6001 (10 µM). Data are shown as the mean ± SEM (n = 4–5/group). * P

    Article Snippet: Tyrphostin AG 1478, Phenylephrine, Apocynin, Peg-Catalase (PG-Cat), Dihydroethidium (DHE), phenanthroline, Phenylmethylsulfonyl fluoride were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: Activity Assay, Incubation, In Situ, Zymography, Fluorescence

    Biochemical properties of HCA. ( A ) Activity–pH relationship curve; ( B ) activity–temperature relationship curve; ( C ) effects of Ca 2+ on the fibrinogen clotting activity; ( D ) effect of PMSF on the clotting time of fibrinogen solution after the addition of HCA. The values shown are the mean values ± SEM (n=3). Abbreviations: HCA, hemocoagulase agkistrodon; PMSF, phenylmethanesulfonyl fluoride; SEM, standard error of mean.

    Journal: Drug Design, Development and Therapy

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities

    doi: 10.2147/DDDT.S159210

    Figure Lengend Snippet: Biochemical properties of HCA. ( A ) Activity–pH relationship curve; ( B ) activity–temperature relationship curve; ( C ) effects of Ca 2+ on the fibrinogen clotting activity; ( D ) effect of PMSF on the clotting time of fibrinogen solution after the addition of HCA. The values shown are the mean values ± SEM (n=3). Abbreviations: HCA, hemocoagulase agkistrodon; PMSF, phenylmethanesulfonyl fluoride; SEM, standard error of mean.

    Article Snippet: HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00.

    Techniques: High Content Screening, Activity Assay, Coagulation

    Trypsin digestion of Vip3Aa using inhibitors to stop the reaction. The reactions were stopped with the addition of irreversible trypsin protease inhibitors or urea. Loading buffer was added either immediately ( a ) or 10 min after addition of the inhibitors or the urea ( b ) and the samples heated and subjected to SDS-PAGE. Lanes 1: molecular weight markers; lanes 2: untreated protoxin; lanes 3: control with no inhibitors; lanes 4: 1 mM PMSF; lanes 5: 0.1 mM TLCK; lanes 6: 1 mM AEBSF; lanes 7: 10 mM E64; lanes 8: 8 M urea. Molecular weight markers are indicated in kDa.

    Journal: Toxins

    Article Title: Insights into the Structure of the Vip3Aa Insecticidal Protein by Protease Digestion Analysis

    doi: 10.3390/toxins9040131

    Figure Lengend Snippet: Trypsin digestion of Vip3Aa using inhibitors to stop the reaction. The reactions were stopped with the addition of irreversible trypsin protease inhibitors or urea. Loading buffer was added either immediately ( a ) or 10 min after addition of the inhibitors or the urea ( b ) and the samples heated and subjected to SDS-PAGE. Lanes 1: molecular weight markers; lanes 2: untreated protoxin; lanes 3: control with no inhibitors; lanes 4: 1 mM PMSF; lanes 5: 0.1 mM TLCK; lanes 6: 1 mM AEBSF; lanes 7: 10 mM E64; lanes 8: 8 M urea. Molecular weight markers are indicated in kDa.

    Article Snippet: The irreversible inhibitors used to stop the trypsin action, were: PMSF (phenylmethanesulfonyl fluoride, from SIGMA), TLCK (Nα-tosyl-l -lysine chloromethyl ketone hydrochloride, from SIGMA), AEBSF protease inhibitor (from ThermoFisher, Waltham, MA, USA), and E64 (trans-epoxysuccinyl-l -leucylamido (4-guanidino) butane, from SIGMA).

    Techniques: SDS Page, Molecular Weight

    Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Link between light and fatty acid synthesis: Thioredoxin-linked reductive activation of plastidic acetyl-CoA carboxylase

    doi:

    Figure Lengend Snippet: Effects of thiol reagents on the ACCase activity. ( A ) ACCase activity was measured in the indicated concentration of a thiol reagent as described in Materials and Methods . Plastidic ACCase from pea, continuous lines; cytosolic ACCase from pea, dashed line. β-SH, 2-mercaptoethanol; GSH, reduced form of glutathione. Lipoic acid was dissolved into 50 mM Tricine⋅KOH (pH 8) containing 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid at 20 mM, and 0–7 μl of 20 mM solution was added to 13-μl reaction mixture. The activity without any thiol reagent was 11 nmol/20 min per mg of protein for pea plastidic ACCase and 210 nmol/20 min per mg of protein for pea cytosolic ACCase. The activity is relative to that without a thiol reagent, taken to be 1. Results are the means of duplicate determinations that differed less than 10%. ( B ) The effects of preincubation of plastidic ACCase with DTT on activity were examined. Changes with time are shown. After the enzyme was incubated at 30°C for 10 min with 2 mM DTT, the reaction was started by the addition of buffer, ATP, MgCl 2 , KCl, acetyl-CoA, and NaH 14 CO 3 . The final concentrations of these reagents were the same as above. The final concentration of DTT was 1 mM.

    Article Snippet: The ammonium sulfate precipitates of the plastidic or the cytosolic ACCase fraction were dissolved in 50–100 μl of a 50 mM Tricine⋅KOH (pH 8) buffer containing 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamide, and 5 mM ɛ-amino-n-caproic acid, and desalted by centrifugation in a gel filtration column (Biospin 6, or Biospin 30, Bio-Rad).

    Techniques: Activity Assay, Concentration Assay, Incubation

    Effect of CdTe QDs of different sizes on autophagy in yeast at different time points. Notes: Cells transformed with a plasmid encoding GFP-Atg8 were treated with ( A ) 50 nmol/L CdTe QDs or 100 ng/mL rapamycin or ( B ) phenylmethanesulfonyl fluoride (PMSF), which was added at 0 hour and 16 hours to reach a final concentration of 2 mmol/L, and photographed at 4 hours, 8 hours, 16 hours, and 42 hours using fluorescence microscopy. Scale bars: 5 µm. Abbreviations: CdTe QDs, cadmium telluride quantum dots; G-CdTe, green-emitting CdTe; O-CdTe, orange-emitting CdTe.

    Journal: International Journal of Nanomedicine

    Article Title: Inhibition of autophagy contributes to the toxicity of cadmium telluride quantum dots in Saccharomyces cerevisiae

    doi: 10.2147/IJN.S108636

    Figure Lengend Snippet: Effect of CdTe QDs of different sizes on autophagy in yeast at different time points. Notes: Cells transformed with a plasmid encoding GFP-Atg8 were treated with ( A ) 50 nmol/L CdTe QDs or 100 ng/mL rapamycin or ( B ) phenylmethanesulfonyl fluoride (PMSF), which was added at 0 hour and 16 hours to reach a final concentration of 2 mmol/L, and photographed at 4 hours, 8 hours, 16 hours, and 42 hours using fluorescence microscopy. Scale bars: 5 µm. Abbreviations: CdTe QDs, cadmium telluride quantum dots; G-CdTe, green-emitting CdTe; O-CdTe, orange-emitting CdTe.

    Article Snippet: Phenylmethanesulfonyl fluoride was obtained from Amresco (Solon, OH, USA).

    Techniques: Transformation Assay, Plasmid Preparation, Concentration Assay, Fluorescence, Microscopy

    Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF phenylmethylsulfonyl fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown

    Journal: Malaria Journal

    Article Title: The malaria parasite Plasmodium falciparum in red blood cells selectively takes up serum proteins that affect host pathogenicity

    doi: 10.1186/s12936-020-03229-1

    Figure Lengend Snippet: Coagulation occurred after parasite egress from iRBCs. a – c (i) Western blotting analysis of coagulation. a “Med” and “Sup” indicate PDS- or NCS-containing medium and PDS- or NCS-cultured supernatant, respectively. b After the addition of protease inhibitors ( PMSF phenylmethylsulfonyl fluoride, DE dabigatran etexilate). c “Lysate” indicates the isolated parasite lysate prepared using the precipitation method. The isolated parasite lysate prepared from iRBCs cultured in NCS-containing medium was added to plasma. (i) The absence of fibrinogen indicated coagulation, save for in the isolated parasite lysate (lanes 3 and 4). (ii) Visual confirmation of coagulation. Tubes, shown in lanes 3, 4, 5, and 6 in (i), containing 100 µL of the reaction mixture were rotated at a 90° angle onto their sides after completion of the coagulation reaction. All experiments were performed in triplicate and representative data are shown

    Article Snippet: Phenylmethylsulfonyl fluoride

    Techniques: Coagulation, Western Blot, Cell Culture, Isolation