protease inhibitor cocktail Roche Search Results


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  • 99
    Thermo Fisher protease inhibitor cocktails
    Protease Inhibitor Cocktails, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protease inhibitor cocktail roche
    Protease Inhibitor Cocktail Roche, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 13124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche protease inhibitor cocktail roche
    Protease Inhibitor Cocktail Roche, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche roche protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Roche Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 1126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche complete protease inhibitor cocktail roche
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Complete Protease Inhibitor Cocktail Roche, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche protease inhibitor cocktail mini complete roche
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Protease Inhibitor Cocktail Mini Complete Roche, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche mini protease inhibitor cocktail roche 11836153001
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Mini Protease Inhibitor Cocktail Roche 11836153001, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protease inhibitors
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Protease Inhibitors, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 30453 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Protease Inhibitor Cocktail, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 109921 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche 1×roche complete mini protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    1×Roche Complete Mini Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche 1x roche protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    1x Roche Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche easypack roche protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    Easypack Roche Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche 2x roche protease inhibitor cocktail
    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma <t>protease</t> <t>cocktail.</t> a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease <t>inhibitor;</t> Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 <t>Roche</t> Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months
    2x Roche Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche roche halt protease inhibitor cocktails
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
    Roche Halt Protease Inhibitor Cocktails, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc protease inhibitor cocktail
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
    Protease Inhibitor Cocktail, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore complete protease inhibitor cocktail
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
    Complete Protease Inhibitor Cocktail, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1400 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche protease inhibitor cocktail roche complete minitablets
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
    Protease Inhibitor Cocktail Roche Complete Minitablets, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioVision cocktail
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Boehringer Mannheim protease inhibitor cocktail
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Roche mammalian protease inhibitors
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Roche ultra protease inhibitors
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Millipore complete mini protease inhibitor cocktail
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Roche edta free protease inhibitor cocktail cpi tablets roche
    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 <t>inhibitor.</t> H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% <t>Roche/Halt</t> <t>protease</t> inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.
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    Image Search Results


    Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma protease cocktail. a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease inhibitor; Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 Roche Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months

    Journal: BMC Nephrology

    Article Title: Strategy and rationale for urine collection protocols employed in the NEPTUNE study

    doi: 10.1186/s12882-015-0185-3

    Figure Lengend Snippet: Western blot of exosome pellets. Exosomal proteins of interest including podocalyxin (PODXL), fibrocystin (FIBRO) and polycystin 1 (PC1) were examined by Western blot after storage and compared with freshly-isolated exosomes. These proteins were best preserved by the Sigma protease cocktail. a Polycystin 1 detection by Western blot was stable for 1 week at room temperature after exosomes were isolated and stored in 0.01 % sodium azide ( n = 7). However, polycystin 1 was not detected well from exosomes isolated after urine was frozen.at -80C probably because of precipitation issues and unlikely due to loss of protease activity. Effects of preservatives on Western blot of exosome pellet proteins PODXL, scramblase (PLSCR1), FIBRO, and smoothened (SMO). b Lane 1, Fresh, no preservatives; Lane 2, Fresh, no preservatives; Lane 3, −80 °C Frozen/Thawed; Lane 4, −80 °C Frozen/Thawed. c Comparison of varied amounts of protease inhibitors on exosome protein detection by Western blot. Initial assessment and following 6 months storage at −80 °C. Lane 1, 4.8 μL. Sigma protease inhibitor; Lane 2, 4.0 μL Sigma protease inhibitor; Lane 3, 1:100 Roche Complete tablet; Lane 4, −80 °C Frozen/Thawed; Lane 5, Fresh, no preservatives. d Exosomal Podocalyxin fared well at RT and at 4 °C. Sodium azide did not affect their survival. e Analysis of exosomes extracted from frozen (−80 °C) raw urine stored for 12 months

    Article Snippet: There was substantially better detection of the podocyte marker of interest PODXL in cellular pellets stored with protease inhibitors, and again found that Sigma protease inhibitor product may in fact be superior to the more expensive Roche protease inhibitor cocktail (Additional file : Figure S1).

    Techniques: Western Blot, Isolation, Activity Assay, Protease Inhibitor

    Miro1 K572R mutation does not affect Miro1 function in mitochondrial motility in neurons Suppression of Miro1 and Miro2 with shRNAs led to an almost complete inhibition of mitochondrial motility; co‐expression of shRNAs with shRNA‐resistant WT Miro1 or K572R mutant both efficiently rescued mitochondrial motility (left panel: fraction of time in motion, middle panel: relative mitochondrial velocity). Data are presented as a Tukey boxplot. **** P

    Journal: The EMBO Journal

    Article Title: Miro proteins prime mitochondria for Parkin translocation and mitophagy

    doi: 10.15252/embj.201899384

    Figure Lengend Snippet: Miro1 K572R mutation does not affect Miro1 function in mitochondrial motility in neurons Suppression of Miro1 and Miro2 with shRNAs led to an almost complete inhibition of mitochondrial motility; co‐expression of shRNAs with shRNA‐resistant WT Miro1 or K572R mutant both efficiently rescued mitochondrial motility (left panel: fraction of time in motion, middle panel: relative mitochondrial velocity). Data are presented as a Tukey boxplot. **** P

    Article Snippet: Media and reagents Reagents were from the following manufacturers: 35‐mm glass‐bottomed dishes (MatTek, P35G‐1.0‐14‐C), CELLview™ dishes (Greiner Bio‐One 627 871), poly‐L‐lysine (Sigma‐Aldrich, P6282), collagen IV (Sigma‐Aldrich, C5533), penicillin, streptomycin and amphotericin B (Invitrogen, 15240‐096), gentamicin (Krka, d. d., Novo Mesto, Slovakia), RPMI‐1640 (Life Technologies, A1049101), DMEM (Life Technologies, 31966021), Neurobasal™ A (Life Technologies, 10888022 and 12349015 (without phenol red)), HBSS 1640 (Life Technologies, 14025050), BME (Life Technologies 41010‐109), Opti‐MEM I (Life Technologies, 31985‐047), B‐27® Supplement (50X, serum free, Life Technologies, 17504044), foetal bovine serum (Life Technologies, 10270106), horse serum (Life Technologies, 26050088), GlutaMAX™‐I (Life Technologies, 3505038) Lipofectamine® 2000 (Invitrogen, 11668‐019), Metafectene® (Biontex Laboratories GmbH, T040), DMSO (Sigma‐Aldrich, 472301), FCCP (Tocris, 0453), G418 (Sigma‐Aldrich, A1720), MG132 (Tocris, 1748), protease inhibitor cocktail cOmplete (Roche, 04693116001), protein G‐sepharose 4B beads (Invitrogen, 101243), DC Protein Assay reagent (Bio‐Rad, 500‐0111), Pierce Lane Marker Reducing sample buffer (Thermo Fisher Scientific, 39000).

    Techniques: Mutagenesis, Inhibition, Expressing, shRNA

    Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 inhibitor. H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% Roche/Halt protease inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.

    Journal: Journal of Cancer

    Article Title: Modification of proteolytic activity matrix analysis (PrAMA) to measure ADAM10 and ADAM17 sheddase activities in cell and tissue lysates

    doi: 10.7150/jca.20779

    Figure Lengend Snippet: Modified PrAMA detects decreases of ADAM10sa in human-cell and -tissue lysates exposed to ADAM10 inhibitor. H441-cell ( A ) and T2495-tissue ( B ) lysates (2 μg) were supplemented with 0.5% Roche/Halt protease inhibitors, vehicle (Control) or GI254023X (1 μM), and PEPDAB substrates (10 μM), and incubated for 4 h. The developing fluorescence was measured hourly. The presented data are pM means of duplicate measurements ± SD of processed substrates, and are representative of ten experiments performed. The GI254023X-mediated inhibitions of PEPDAB005, 010 and/or 022 processing with the lysates are significant (H441: p=0.017, 0.0041 and 0.0058; T2495: p=0.05, 0.04 and 0.08, respectively). The substrate processing data were analyzed using the systematically increased Syntherror/Sigmathreshold parameters from 0.5/0.0 to 0.5/2.0. The obtained data at 4 h incubation are presented as PrAMA ADAM10sa AU of H441-cell ( C ) and T2495-tissue ( D ) lysates. PrAMA standard errors were 1.2% to 9%. Decreases of ADAM10sa in H441-cell ( E ) and T2495-tissue ( F ) lysates are presented as % of the specific enzyme activities in the control lysates relative to the GI254023X treated lysates.

    Article Snippet: These data show that Tris-based assay buffer supplemented with 0.5% combined Roche/Halt protease-inhibitor cocktails strongly suppress non-MP proteases without affecting recombinant or natural ADAM10sa and ADAM17sa.

    Techniques: Modification, Incubation, Fluorescence