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  • 95
    Millipore dimethylammonio 1 propanesulfonate hydrate
    Dimethylammonio 1 Propanesulfonate Hydrate, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 2 article reviews
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    dimethylammonio 1 propanesulfonate hydrate - by Bioz Stars, 2020-02
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    77
    Thermo Fisher zoom chaps
    Zoom Chaps, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 77 stars, based on 2 article reviews
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    79
    Avantor chaps
    Chaps, supplied by Avantor, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 79 stars, based on 5 article reviews
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    85
    Merck KGaA chaps
    Chaps, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 6 article reviews
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    80
    GE Healthcare chaps
    Chaps, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 80/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 80 stars, based on 16 article reviews
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    79
    BioShop chaps
    Chaps, supplied by BioShop, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 79 stars, based on 4 article reviews
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    99
    Millipore chaps buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 360 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc chaps buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps Buffer, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 58 article reviews
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    chaps buffer - by Bioz Stars, 2020-02
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    88
    Applichem chaps buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps Buffer, supplied by Applichem, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore chaps
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Cell Signaling Technology Inc 1x chaps buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    1x Chaps Buffer, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Valiant zwitterionic surfactant chaps
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Zwitterionic Surfactant Chaps, supplied by Valiant, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    FIVEphoton Biochemicals chaps lysis buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps Lysis Buffer, supplied by FIVEphoton Biochemicals, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chaps lysis buffer - by Bioz Stars, 2020-02
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    99
    Protein Simple Inc bicine chaps buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Bicine Chaps Buffer, supplied by Protein Simple Inc, used in various techniques. Bioz Stars score: 99/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bicine chaps buffer - by Bioz Stars, 2020-02
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    93
    Merck KGaA chaps lysis buffer
    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the <t>CHAPS</t> buffer and <t>immunoprecipitation</t> with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.
    Chaps Lysis Buffer, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore chaps lysis buffer
    Surface marker expression, SA-β-gal staining and telomerase activity in HBCEC . A . Determination of the percentage of cell surface marker expression in HBCEC at different ages. Expression of the surface marker proteins CD24, CD44, CD227 was maintained during long term culture of HBCEC. Whereas CD24 and CD44 were similarly expressed after 176d and 462d, CD227 increased from 52% to 88% in HBCEC 462d. The flow cytometry measurements varied by about 8%. B . SA-β-gal staining of primary HBCEC and HMEC cultures. Staining for SA-β-gal of a HBCEC population after 722d in culture revealed little if any positive cell. Normal HMEC in passage 16, however, displayed already predominantly enlarged senescent cells after 32d, demonstrated by the dark-green stain (bar = 200 μm). C . Telomerase (TRAP-)assay of primary cultures from breast cancer biopsies. Telomerase activity was analyzed according to the Telomeric Repeat Amplification Protocol (TRAP). HBCEC populations demonstrated telomerase activity independent of the age of the culture and the harvest method. The human embryonic kidney (HEK) 293T cell line was used as a positive control and <t>1×</t> <t>CHAPS</t> buffer served as a negative control. Quantification was performed using densitometric analysis.
    Chaps Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 453 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Beyotime chaps lysis buffer
    Surface marker expression, SA-β-gal staining and telomerase activity in HBCEC . A . Determination of the percentage of cell surface marker expression in HBCEC at different ages. Expression of the surface marker proteins CD24, CD44, CD227 was maintained during long term culture of HBCEC. Whereas CD24 and CD44 were similarly expressed after 176d and 462d, CD227 increased from 52% to 88% in HBCEC 462d. The flow cytometry measurements varied by about 8%. B . SA-β-gal staining of primary HBCEC and HMEC cultures. Staining for SA-β-gal of a HBCEC population after 722d in culture revealed little if any positive cell. Normal HMEC in passage 16, however, displayed already predominantly enlarged senescent cells after 32d, demonstrated by the dark-green stain (bar = 200 μm). C . Telomerase (TRAP-)assay of primary cultures from breast cancer biopsies. Telomerase activity was analyzed according to the Telomeric Repeat Amplification Protocol (TRAP). HBCEC populations demonstrated telomerase activity independent of the age of the culture and the harvest method. The human embryonic kidney (HEK) 293T cell line was used as a positive control and <t>1×</t> <t>CHAPS</t> buffer served as a negative control. Quantification was performed using densitometric analysis.
    Chaps Lysis Buffer, supplied by Beyotime, used in various techniques. Bioz Stars score: 89/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Boston BioProducts chaps lysis buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Lysis Buffer, supplied by Boston BioProducts, used in various techniques. Bioz Stars score: 89/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Roche chaps zwitterionic detergent
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Zwitterionic Detergent, supplied by Roche, used in various techniques. Bioz Stars score: 82/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cell Signaling Technology Inc chaps cell extract buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Cell Extract Buffer, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Roche chaps buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher chaps buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Anatrace chaps anagrade
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Anagrade, supplied by Anatrace, used in various techniques. Bioz Stars score: 75/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher chaps anagrade
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Anagrade, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    GE Healthcare cholamidopropyl chaps
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Cholamidopropyl Chaps, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad chaps buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Chaps Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore 1x chaps lysis buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    1x Chaps Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GE Healthcare dimethylammonio 1 propanesulfonate chaps
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Dimethylammonio 1 Propanesulfonate Chaps, supplied by GE Healthcare, 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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    Millipore trapeze chaps lysis buffer
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
    Trapeze Chaps Lysis Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    AG Scientific cholamidopropyl dimethylammonio 1 propane chaps
    Phenobarbital and <t>CITCO</t> treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL <t>CHAPS</t> lysis buffer. Lysates were
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    N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the CHAPS buffer and immunoprecipitation with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.

    Journal: Oncotarget

    Article Title: The anti-HER3 (ErbB3) therapeutic antibody 9F7-F11 induces HER3 ubiquitination and degradation in tumors through JNK1/2- dependent ITCH/AIP4 activation

    doi: 10.18632/oncotarget.9455

    Figure Lengend Snippet: N4BP1 overexpression inhibits 9F7-F11-mediated HER3 ubiquitination and degradation induced by ITCH, and promotes HER3 protein stabilization in BxPc3 cells ( A ) Cells were co-transfected with the Myc-ITCHwt, HA-Ub and/or GFP-N4BP1 plasmids for 24 hr. Transfected cells were then incubated with 20 μM MG132 for 5 hr before addition of 50 μg/mL 9F7-F11 for 3 hr. After cells lysis with the CHAPS buffer and immunoprecipitation with the HER3 Ab, the ubiquitination status was analyzed by western blotting using an anti-HA antibody. HER3 and ITCH were detected using specific antibodies. ( B ) Cells were transfected with increasing doses of the V5-N4BP1 plasmid for 24 hr, and then incubated with 50 μg/mL 9F7-F11 or with 100 ng/mL NRG-1β for 3 hr. HER3 expression was assessed in whole cell lysates by western blotting. The V5-HRP antibody was used to detect N4BP1, and β-actin was the loading control. The signal intensity (SI) of the different bands was quantified with ImageJ. WCL, whole cell lysates.

    Article Snippet: Cell lysis and immunoprecipitation Cells were lysed in CHAPS buffer (Sigma-Aldrich) containing the protease inhibitor cocktail V (Calbiochem, Billerica, MA) and the phosphatase inhibitor cocktail II (Sigma-Aldrich).

    Techniques: Over Expression, Transfection, Incubation, Lysis, Immunoprecipitation, Western Blot, Plasmid Preparation, Expressing

    Surface marker expression, SA-β-gal staining and telomerase activity in HBCEC . A . Determination of the percentage of cell surface marker expression in HBCEC at different ages. Expression of the surface marker proteins CD24, CD44, CD227 was maintained during long term culture of HBCEC. Whereas CD24 and CD44 were similarly expressed after 176d and 462d, CD227 increased from 52% to 88% in HBCEC 462d. The flow cytometry measurements varied by about 8%. B . SA-β-gal staining of primary HBCEC and HMEC cultures. Staining for SA-β-gal of a HBCEC population after 722d in culture revealed little if any positive cell. Normal HMEC in passage 16, however, displayed already predominantly enlarged senescent cells after 32d, demonstrated by the dark-green stain (bar = 200 μm). C . Telomerase (TRAP-)assay of primary cultures from breast cancer biopsies. Telomerase activity was analyzed according to the Telomeric Repeat Amplification Protocol (TRAP). HBCEC populations demonstrated telomerase activity independent of the age of the culture and the harvest method. The human embryonic kidney (HEK) 293T cell line was used as a positive control and 1× CHAPS buffer served as a negative control. Quantification was performed using densitometric analysis.

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Characterization of human breast cancer epithelial cells (HBCEC) derived from long term cultured biopsies

    doi: 10.1186/1756-9966-28-127

    Figure Lengend Snippet: Surface marker expression, SA-β-gal staining and telomerase activity in HBCEC . A . Determination of the percentage of cell surface marker expression in HBCEC at different ages. Expression of the surface marker proteins CD24, CD44, CD227 was maintained during long term culture of HBCEC. Whereas CD24 and CD44 were similarly expressed after 176d and 462d, CD227 increased from 52% to 88% in HBCEC 462d. The flow cytometry measurements varied by about 8%. B . SA-β-gal staining of primary HBCEC and HMEC cultures. Staining for SA-β-gal of a HBCEC population after 722d in culture revealed little if any positive cell. Normal HMEC in passage 16, however, displayed already predominantly enlarged senescent cells after 32d, demonstrated by the dark-green stain (bar = 200 μm). C . Telomerase (TRAP-)assay of primary cultures from breast cancer biopsies. Telomerase activity was analyzed according to the Telomeric Repeat Amplification Protocol (TRAP). HBCEC populations demonstrated telomerase activity independent of the age of the culture and the harvest method. The human embryonic kidney (HEK) 293T cell line was used as a positive control and 1× CHAPS buffer served as a negative control. Quantification was performed using densitometric analysis.

    Article Snippet: Briefly, HBCEC and 293T control cells were washed with ice-cold PBS and homogenized in 100 μl ice-cold 1× CHAPS lysis buffer (Chemicon).

    Techniques: Marker, Expressing, Staining, Activity Assay, Flow Cytometry, Cytometry, TRAP Assay, Amplification, Positive Control, Negative Control

    Phenobarbital and CITCO treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL CHAPS lysis buffer. Lysates were

    Journal: Archives of biochemistry and biophysics

    Article Title: Regulation of the human cathepsin E gene by the constitutive androstane receptor

    doi: 10.1016/j.abb.2007.08.001

    Figure Lengend Snippet: Phenobarbital and CITCO treatment of human hepatocytes results in increased cathepsin E enzyme activity. Cultures of primary human hepatocytes, control and those treated with PB or CITCO, were harvested in 150 μL CHAPS lysis buffer. Lysates were

    Article Snippet: Five day cultures of primary human hepatocytes, control or treated for 24 h with 500 μM PB or 5 μM CITCO, were lysed in 150 μL of CHAPS lysis buffer (Boston Bioproducts, Worcester, MA).

    Techniques: Activity Assay, Lysis