protease inhibitor cocktail mini complete roche  (Roche)

 
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    Structured Review

    Roche protease inhibitor cocktail mini complete roche
    Protease Inhibitor Cocktail Mini Complete Roche, supplied by Roche, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease inhibitor cocktail mini complete roche/product/Roche
    Average 89 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail mini complete roche - by Bioz Stars, 2020-04
    89/100 stars

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    Related Articles

    Transfection:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: After 48 h of transfection, the cells were lysed by the addition of Cell Culture Lysis Buffer (Promega, USA), followed by incubation at room temperature for 5 min. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Vero cells were transiently transfected with pSGFlag-eIF3L; after 24 h, the cells were infected with YFV strain 17D at an M.O.I. of 5 and incubated in DMEM medium without FBS for 2 h. The infected cells were washed with PBS and reincubated in DMEM medium supplemented with 1% FBS for 48 h in the presence of 5% CO2. .. The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®).

    Luciferase:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: Overexpression experiments and luciferase activity assays ... The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    In Vitro:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: In vitro binding assay ... After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C).

    Positive Control:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The empty vector and untransfected cells were used as the negative controls; RNAi against NS1 YFV [ ] was used as a positive control. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Binding Assay:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: In vitro binding assay ... After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C).

    Isolation:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cell supernatants were harvested, and RNA was isolated using the viral RNA isolation kit (HiSpeed Plasmid Midi Kit, Qiagen). .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    Plasmid Preparation:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The empty vector and untransfected cells were used as the negative controls; RNAi against NS1 YFV [ ] was used as a positive control. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cell supernatants were harvested, and RNA was isolated using the viral RNA isolation kit (HiSpeed Plasmid Midi Kit, Qiagen). .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    Infection:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication. .. At 24 h after transfection, the cells were counted and infected with YFV 17D at an M.O.I. of 1 for 2 h (37°C).

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Vero cells were transiently transfected with pSGFlag-eIF3L; after 24 h, the cells were infected with YFV strain 17D at an M.O.I. of 5 and incubated in DMEM medium without FBS for 2 h. The infected cells were washed with PBS and reincubated in DMEM medium supplemented with 1% FBS for 48 h in the presence of 5% CO2. .. The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®).

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: Knock-down experiments using shRNA and infection assays ... The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    Real-time Polymerase Chain Reaction:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The viral RNA copy number was determined using real-time PCR analysis, as described above. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    Immunoprecipitation:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®). .. Equal amounts of protein extracts were subjected to immunoprecipitation for 3 h (4°C) using anti-Flag M2 Affinity gel beads.

    Incubation:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: After 48 h of transfection, the cells were lysed by the addition of Cell Culture Lysis Buffer (Promega, USA), followed by incubation at room temperature for 5 min. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Vero cells were transiently transfected with pSGFlag-eIF3L; after 24 h, the cells were infected with YFV strain 17D at an M.O.I. of 5 and incubated in DMEM medium without FBS for 2 h. The infected cells were washed with PBS and reincubated in DMEM medium supplemented with 1% FBS for 48 h in the presence of 5% CO2. .. The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®).

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Knock-down experiments using shRNA and infection assays HEK-293T cells were infected with pseudotyped lentiviral particles at an M.O.I. of 0.1 for 2 h. The infected cells were washed with PBS and incubated in DMEM medium supplemented with 1% FCS for 72 h in the presence of 5% CO2 . .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    shRNA:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: Knock-down experiments using shRNA and infection assays ... The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above.

    Activity Assay:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: Paragraph title: Overexpression experiments and luciferase activity assays ... The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Cell Culture:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: After 48 h of transfection, the cells were lysed by the addition of Cell Culture Lysis Buffer (Promega, USA), followed by incubation at room temperature for 5 min. .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication.

    Protease Inhibitor:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication. .. For these assays, Vero cells were transfected with the pCDNAFlag-eIF3L plasmid or a control using the Turbofect transfection reagent (ThermoScientific) at a ratio of 1:3, essentially as described by the manufacturer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®). .. Equal amounts of protein extracts were subjected to immunoprecipitation for 3 h (4°C) using anti-Flag M2 Affinity gel beads.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. .. Plaque reduction assay Vero cells were plated in 6-well microplates (4 × 105 cells/well) and transfected with pCDNAFlag-eIF3L plasmid using the Lipofectamine™ 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

    Western Blot:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication. .. For these assays, Vero cells were transfected with the pCDNAFlag-eIF3L plasmid or a control using the Turbofect transfection reagent (ThermoScientific) at a ratio of 1:3, essentially as described by the manufacturer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®). .. The antibody-protein complexes were then resolved by SDS-PAGE, and the YFV NS5 protein was identified by western blotting using an anti-YFNS5 (1:4000) antibody, a kind gift of Charles M. Rice, Rockefeller University.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. .. Plaque reduction assay Vero cells were plated in 6-well microplates (4 × 105 cells/well) and transfected with pCDNAFlag-eIF3L plasmid using the Lipofectamine™ 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

    Lysis:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication. .. For these assays, Vero cells were transfected with the pCDNAFlag-eIF3L plasmid or a control using the Turbofect transfection reagent (ThermoScientific) at a ratio of 1:3, essentially as described by the manufacturer.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®). .. Equal amounts of protein extracts were subjected to immunoprecipitation for 3 h (4°C) using anti-Flag M2 Affinity gel beads.

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. .. Plaque reduction assay Vero cells were plated in 6-well microplates (4 × 105 cells/well) and transfected with pCDNAFlag-eIF3L plasmid using the Lipofectamine™ 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

    Over Expression:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. The cells were also harvested in lysis buffer (50 mM Tris–HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM dithiothreitol, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®) for a western blot analysis, as described above. eIF3L overexpression was also performed in Vero cells to better examine the alterations in the kinetics of viral replication. .. For these assays, Vero cells were transfected with the pCDNAFlag-eIF3L plasmid or a control using the Turbofect transfection reagent (ThermoScientific) at a ratio of 1:3, essentially as described by the manufacturer.

    SDS Page:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: The cells were harvested in lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.1% NP-40, 5% glycerol, 1 mM sodium fluoride, 0.1 mM sodium orthovanadate, 10 mM β-glycerolphosphate, and protease inhibitor cocktail Mini complete Roche®). .. The antibody-protein complexes were then resolved by SDS-PAGE, and the YFV NS5 protein was identified by western blotting using an anti-YFNS5 (1:4000) antibody, a kind gift of Charles M. Rice, Rockefeller University.

    Derivative Assay:

    Article Title: The eukaryotic translation initiation factor 3 subunit L protein interacts with Flavivirus NS5 and may modulate yellow fever virus replication
    Article Snippet: .. After four washes with 1 ml phosphate-buffered saline (PBS) and a final wash with lysis buffer (50 mM Tris–HCl [pH 7.5], 150 mM NaCl, 0.5% NP-40, 1 mM sodium fluoride, 0.2 mM sodium orthovanadate, 10 mM β-glycerophosphate, and protease inhibitor cocktail Mini complete Roche®), 600 μg of extract derived from HeLa cells transfected with the eIF3L-pSGFlag plasmid was added and incubated for 2 h (4°C). .. The unbound fractions were collected, and the beads were washed four times with 1 ml of lysis buffer.

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  • 99
    Roche complete mini protease inhibitor cocktail
    Expression of conventional breast cancer biomarkers and selected multidrug resistance genes. Cell lysates were prepared in radioimmunoprecipitation assay buffer supplemented with <t>cOmplete</t> Mini protease inhibitor and PhosSTOP phosphatase inhibitor. Quantities (10–50 μg) of total protein were run on a 10 % gel (MDR1), 4–20 % precast gels (epidermal growth factor receptor [EGFR], oestrogen receptor [ER], progesterone receptor [PR], type II topoisomerase [TOPO IIα]) and Any kD Mini-PROTEAN TGX Precast Protein Gels (human epidermal growth factor receptor 2 [HER2], HER3), transferred onto polyvinylidene fluoride membrane and developed using chemiluminescence substrate. Nat native, Epi-R epirubicin-resistant, GAPDH glyceraldehyde-3-phosphate dehydrogenase
    Complete Mini Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 351 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete mini protease inhibitor cocktail/product/Roche
    Average 99 stars, based on 351 article reviews
    Price from $9.99 to $1999.99
    complete mini protease inhibitor cocktail - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Roche complete mini edta free protease inhibitor cocktail
    Apocatalase polypeptide proteolysis is induced in late exponential growth phase E. faecalis cells. A , lysates from strain OG1RF cells grown in heme-free medium until early exponential growth phase ( sample A ) or late exponential growth phase ( sample B ) and lysates from cells grown in heme-supplemented medium until late exponential growth phase ( sample C ) were analyzed individually or as mixtures as indicated. The samples were incubated at 37 °C, and the presence of KatA polypeptide in aliquots removed at the indicated time points was determined by immuno-blot. B , proteolysis of KatA in sample A mixed with sample B in the presence of 1 m m <t>PMSF</t> and/or <t>EDTA</t> or Complete EDTA-free protease inhibitor cocktail. The reference, sample A + B without any additions, is indicated by Ctrl .
    Complete Mini Edta Free Protease Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete mini edta free protease inhibitor cocktail/product/Roche
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    complete mini edta free protease inhibitor cocktail - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    Expression of conventional breast cancer biomarkers and selected multidrug resistance genes. Cell lysates were prepared in radioimmunoprecipitation assay buffer supplemented with cOmplete Mini protease inhibitor and PhosSTOP phosphatase inhibitor. Quantities (10–50 μg) of total protein were run on a 10 % gel (MDR1), 4–20 % precast gels (epidermal growth factor receptor [EGFR], oestrogen receptor [ER], progesterone receptor [PR], type II topoisomerase [TOPO IIα]) and Any kD Mini-PROTEAN TGX Precast Protein Gels (human epidermal growth factor receptor 2 [HER2], HER3), transferred onto polyvinylidene fluoride membrane and developed using chemiluminescence substrate. Nat native, Epi-R epirubicin-resistant, GAPDH glyceraldehyde-3-phosphate dehydrogenase

    Journal: Breast Cancer Research : BCR

    Article Title: Downregulation of histone H2A and H2B pathways is associated with anthracycline sensitivity in breast cancer

    doi: 10.1186/s13058-016-0676-6

    Figure Lengend Snippet: Expression of conventional breast cancer biomarkers and selected multidrug resistance genes. Cell lysates were prepared in radioimmunoprecipitation assay buffer supplemented with cOmplete Mini protease inhibitor and PhosSTOP phosphatase inhibitor. Quantities (10–50 μg) of total protein were run on a 10 % gel (MDR1), 4–20 % precast gels (epidermal growth factor receptor [EGFR], oestrogen receptor [ER], progesterone receptor [PR], type II topoisomerase [TOPO IIα]) and Any kD Mini-PROTEAN TGX Precast Protein Gels (human epidermal growth factor receptor 2 [HER2], HER3), transferred onto polyvinylidene fluoride membrane and developed using chemiluminescence substrate. Nat native, Epi-R epirubicin-resistant, GAPDH glyceraldehyde-3-phosphate dehydrogenase

    Article Snippet: WCL were prepared by adding equal volumes of 2× RIPA buffer supplemented with 25 U of Benzonase Nuclease (Sigma-Aldrich) and cOmplete Mini Protease Inhibitor Cocktail (Roche), incubating on ice for 30 minutes and sonicating for 15 minutes with 30-second on-off intervals.

    Techniques: Expressing, Radio Immunoprecipitation, Protease Inhibitor

    Giardia duodenalis -induced increase of epithelial sugar uptake is mediated by proteases. (A) Addition of protease inhibitor cocktails, complete mini (CM) decreased G. duodenalis -enhanced sugar uptake. CM alone (CM control) had no effect on baseline enterocytic

    Journal:

    Article Title: SGLT-1-mediated glucose uptake protects human intestinal epithelial cells against Giardia duodenalis-induced apoptosis

    doi: 10.1016/j.ijpara.2007.12.004

    Figure Lengend Snippet: Giardia duodenalis -induced increase of epithelial sugar uptake is mediated by proteases. (A) Addition of protease inhibitor cocktails, complete mini (CM) decreased G. duodenalis -enhanced sugar uptake. CM alone (CM control) had no effect on baseline enterocytic

    Article Snippet: The protease inhibitor cocktail complete mini® which inhibits serine, cysteine, metalloproteases and calpains, the selective cysteine protease inhibitor E64, and the selective serine protease inhibitor Pefabloc SC were purchased from Roche, Laval, Que., Canada.

    Techniques: Protease Inhibitor

    The effect of TCNAs on the viability of human leukemic HL-60 cell line. HL-60 cells were incubated with DOXY, MINO or COL-3 in concentrations within the range 0.5–100 µg/ml for 24 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Viability was studied using resazurin fluorescence assay and expressed as percent of the control. Results are presented as a mean ± SD of three independent experiments.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: The effect of TCNAs on the viability of human leukemic HL-60 cell line. HL-60 cells were incubated with DOXY, MINO or COL-3 in concentrations within the range 0.5–100 µg/ml for 24 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Viability was studied using resazurin fluorescence assay and expressed as percent of the control. Results are presented as a mean ± SD of three independent experiments.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Incubation, Concentration Assay, Fluorescence

    TCNAs-mediated poly (ADP-ribose) polymerase-1(PARP-1) cleavage. HL-60 cells were treated with DOXY or MINO in final concentrations of 2.5, 10 and 25 µg/ml or COL-3 in final concentrations of 0.75, 1 and 5 µg/ml for 6 and 24 h. Cells incubated with DMSO in a final concentration of 0.2% served as controls for solvent toxicity, cells incubated with complete medium served as controls. The pattern of PARP-1 cleavage was assessed using WB. Actin was used as a control for equal protein loading. Co: control, D: DOXY, M: MINO and C: COL-3.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: TCNAs-mediated poly (ADP-ribose) polymerase-1(PARP-1) cleavage. HL-60 cells were treated with DOXY or MINO in final concentrations of 2.5, 10 and 25 µg/ml or COL-3 in final concentrations of 0.75, 1 and 5 µg/ml for 6 and 24 h. Cells incubated with DMSO in a final concentration of 0.2% served as controls for solvent toxicity, cells incubated with complete medium served as controls. The pattern of PARP-1 cleavage was assessed using WB. Actin was used as a control for equal protein loading. Co: control, D: DOXY, M: MINO and C: COL-3.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Incubation, Concentration Assay, Western Blot

    The effect of Z-VAD-FMK on TCNAs-induced cytotoxicity. HL-60 cells were treated with Z-VAD-FMK in a final concentration of 100 µM for 1 h before treatment with TCNAs for 6 h and 24 h. Cells incubated with DMSO in a final concentration of 0.2% served as controls for solvent toxicity, cells incubated in complete medium served as controls. (A) Total cell death was assessed using resazurin viability assay and calculated as 100% - viability %. (B) Apoptosis was assessed by morphological criteria in Giemsa staining. DMSO and Z-VAD-FMK exerted no cytotoxic effects on HL-60 cells compared to controls incubated with complete medium (data not shown on the graph). Results are presented as mean ± SD of three independent experiments. D: DOXY 25 µg/ml, M: MINO 25 µg/ml, Z: Z-VAD-FMK 100 µM; C: COL-3 5 µg/ml in total cell death experiments (A) and 2.5 µg/ml apoptosis experiments (B).

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: The effect of Z-VAD-FMK on TCNAs-induced cytotoxicity. HL-60 cells were treated with Z-VAD-FMK in a final concentration of 100 µM for 1 h before treatment with TCNAs for 6 h and 24 h. Cells incubated with DMSO in a final concentration of 0.2% served as controls for solvent toxicity, cells incubated in complete medium served as controls. (A) Total cell death was assessed using resazurin viability assay and calculated as 100% - viability %. (B) Apoptosis was assessed by morphological criteria in Giemsa staining. DMSO and Z-VAD-FMK exerted no cytotoxic effects on HL-60 cells compared to controls incubated with complete medium (data not shown on the graph). Results are presented as mean ± SD of three independent experiments. D: DOXY 25 µg/ml, M: MINO 25 µg/ml, Z: Z-VAD-FMK 100 µM; C: COL-3 5 µg/ml in total cell death experiments (A) and 2.5 µg/ml apoptosis experiments (B).

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Concentration Assay, Incubation, Viability Assay, Staining

    The role of mitochondria in TCNAs-induced apoptosis. Cytosolic translocation of cytochrome c (cyt c) and Bcl-2 cleavage were assessed with Western blot. Cells were incubated with DOXY or MINO in final concentrations of 2.5, 10 and 25 µg/ml or COL-3 in final concentrations of 0.75, 1 and 5 µg/ml until 24 h. DMSO (0.2%) treated cells served as controls for solvent toxicity, etoposide (6 µg/ml) treated cells as positive controls for apoptosis and cells incubated in complete medium as controls. Co: control, D: DOXY, M: MINO, C: COL-3 and E: etoposide.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: The role of mitochondria in TCNAs-induced apoptosis. Cytosolic translocation of cytochrome c (cyt c) and Bcl-2 cleavage were assessed with Western blot. Cells were incubated with DOXY or MINO in final concentrations of 2.5, 10 and 25 µg/ml or COL-3 in final concentrations of 0.75, 1 and 5 µg/ml until 24 h. DMSO (0.2%) treated cells served as controls for solvent toxicity, etoposide (6 µg/ml) treated cells as positive controls for apoptosis and cells incubated in complete medium as controls. Co: control, D: DOXY, M: MINO, C: COL-3 and E: etoposide.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Translocation Assay, Western Blot, Incubation

    TCNAs-induced apoptosis. HL-60 cells were incubated with DOXY (A) and MINO (B) in concentrations of 5, 10, 25 and 50 µg/ml or COL-3 (C) in concentrations of 0.75, 1, and 5 µg/ml until 48 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Cells were stained with annexin V and PI for 15 min at room temperature in the dark. Twenty thousand events were acquired and analyzed using flow cytometry. Subpopulations are expressed as percentages of total events. Results are expressed as a mean ± SD of three independent experiments. (D), (E): Examples on flow cytometric analysis of TCNAs-induced apoptosis using annexin V/PI assay. D: DOXY, M: MINO COL: COL-3 and PI: propidium iodide.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: TCNAs-induced apoptosis. HL-60 cells were incubated with DOXY (A) and MINO (B) in concentrations of 5, 10, 25 and 50 µg/ml or COL-3 (C) in concentrations of 0.75, 1, and 5 µg/ml until 48 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Cells were stained with annexin V and PI for 15 min at room temperature in the dark. Twenty thousand events were acquired and analyzed using flow cytometry. Subpopulations are expressed as percentages of total events. Results are expressed as a mean ± SD of three independent experiments. (D), (E): Examples on flow cytometric analysis of TCNAs-induced apoptosis using annexin V/PI assay. D: DOXY, M: MINO COL: COL-3 and PI: propidium iodide.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Incubation, Concentration Assay, Staining, Flow Cytometry, Cytometry

    The effect of TCNAs on DNA. HL-60 cells were incubated with DOXY or MINO in final concentrations of 25 µg/ml or COL-3 in a final concentration of 5 µg/ml for 10 min and 1, 2 and 4 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Cells incubated with etoposide in a final concentration of 6 µg/ml served as positive controls for apoptosis. The TCNAs-induced DNA damage was assessed using SDS-PAGE and immunostaining for the DNA double strand breaks marker γH2AX. Co: control, E: etoposide, D: DOXY, M: MINO and C: COL-3.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: The effect of TCNAs on DNA. HL-60 cells were incubated with DOXY or MINO in final concentrations of 25 µg/ml or COL-3 in a final concentration of 5 µg/ml for 10 min and 1, 2 and 4 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Cells incubated with etoposide in a final concentration of 6 µg/ml served as positive controls for apoptosis. The TCNAs-induced DNA damage was assessed using SDS-PAGE and immunostaining for the DNA double strand breaks marker γH2AX. Co: control, E: etoposide, D: DOXY, M: MINO and C: COL-3.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Incubation, Concentration Assay, SDS Page, Immunostaining, Marker

    The loss of mitochondrial membrane potential (Δψm) in TCNAs-induced apoptosis. Mitochondrial membrane potential was assessed using tetramethylrhodamine methyl ester (TMRM) and flow cytometry. (A) HL-60 cells were incubated with DOXY or MINO in final concentrations of 25 and 50 µg/ml or (B) COL-3 in final concentrations of 0.5, 1, 2.5 and 5 µg/ml for 2, 4, 6 and 24 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Results are expressed as mean ± SD of three independent experiments. (C), (D): Example on flow cytometric analysis of loss of Δψm using TMRM assay. D: DOXY, M: MINO, COL: COL-3.

    Journal: PLoS ONE

    Article Title: Cytotoxic Effects of Tetracycline Analogues (Doxycycline, Minocycline and COL-3) in Acute Myeloid Leukemia HL-60 Cells

    doi: 10.1371/journal.pone.0114457

    Figure Lengend Snippet: The loss of mitochondrial membrane potential (Δψm) in TCNAs-induced apoptosis. Mitochondrial membrane potential was assessed using tetramethylrhodamine methyl ester (TMRM) and flow cytometry. (A) HL-60 cells were incubated with DOXY or MINO in final concentrations of 25 and 50 µg/ml or (B) COL-3 in final concentrations of 0.5, 1, 2.5 and 5 µg/ml for 2, 4, 6 and 24 h. Cells incubated in DMSO in a final concentration of 0.2% were used as controls for solvent toxicity and cells incubated in complete medium served as controls. Results are expressed as mean ± SD of three independent experiments. (C), (D): Example on flow cytometric analysis of loss of Δψm using TMRM assay. D: DOXY, M: MINO, COL: COL-3.

    Article Snippet: The other materials were purchased as follows: Z-VAD-FMK (Bachem AG, Bubendorf, Switzerland); etoposide (Bristol-Myers Squibb, Bromma, Sweden); RPMI 1640 medium, Dulbecco’s phosphate-buffer saline (PBS) and fetal bovine serum (FBS) (Invitrogen AB, Stockholm, Sweden); resazurin (R & D System, Stockholm, Sweden); cOmplete mini protease inhibitor cocktail (Roche AB, Stockholm, Sweden); propidium iodide (Sigma Aldrich) and annexin V (BD, Stockholm, Sweden).

    Techniques: Flow Cytometry, Cytometry, Incubation, Concentration Assay

    Apocatalase polypeptide proteolysis is induced in late exponential growth phase E. faecalis cells. A , lysates from strain OG1RF cells grown in heme-free medium until early exponential growth phase ( sample A ) or late exponential growth phase ( sample B ) and lysates from cells grown in heme-supplemented medium until late exponential growth phase ( sample C ) were analyzed individually or as mixtures as indicated. The samples were incubated at 37 °C, and the presence of KatA polypeptide in aliquots removed at the indicated time points was determined by immuno-blot. B , proteolysis of KatA in sample A mixed with sample B in the presence of 1 m m PMSF and/or EDTA or Complete EDTA-free protease inhibitor cocktail. The reference, sample A + B without any additions, is indicated by Ctrl .

    Journal: The Journal of Biological Chemistry

    Article Title: In Vitro Assembly of Catalase *

    doi: 10.1074/jbc.M114.596148

    Figure Lengend Snippet: Apocatalase polypeptide proteolysis is induced in late exponential growth phase E. faecalis cells. A , lysates from strain OG1RF cells grown in heme-free medium until early exponential growth phase ( sample A ) or late exponential growth phase ( sample B ) and lysates from cells grown in heme-supplemented medium until late exponential growth phase ( sample C ) were analyzed individually or as mixtures as indicated. The samples were incubated at 37 °C, and the presence of KatA polypeptide in aliquots removed at the indicated time points was determined by immuno-blot. B , proteolysis of KatA in sample A mixed with sample B in the presence of 1 m m PMSF and/or EDTA or Complete EDTA-free protease inhibitor cocktail. The reference, sample A + B without any additions, is indicated by Ctrl .

    Article Snippet: To test protection of KatA by protease inhibitors 1 mm PMSF and/or 1 mm EDTA, or 1× Complete mini EDTA-free protease inhibitor cocktail (Roche Applied Science), were added to lysate from late exponential growth phase cells.

    Techniques: Incubation, Protease Inhibitor