mgcl2  (Roche)


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    Structured Review

    Roche mgcl2
    Mgcl2, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 2745 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mgcl2/product/Roche
    Average 95 stars, based on 2745 article reviews
    Price from $9.99 to $1999.99
    mgcl2 - by Bioz Stars, 2020-09
    95/100 stars

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    Related Articles

    Centrifugation:

    Article Title: The FA Core Complex Contains a Homo-dimeric Catalytic Module for the Symmetric Mono-ubiquitination of FANCI-FANCD2
    Article Snippet: .. Cells were collected post-infection by centrifugation (5,000 rpm, 4°C, 15 min), washed in PBS buffer supplemented with 5 mM MgCl2 , and resuspended in lysis buffer (25 mM HEPES NaOH [pH 7.6], 15 mM NaCl, 0.02% Tween 20, 10% glycerol, 1 mM EDTA, 1 mM EGTA, 2 mM MgCl2 , 2 mM 2-BME, 0.4 mM PMSF, and Roche protease inhibitor cocktail). .. The suspension was lysed by Dounce homogenization (40 strokes, tight pestle), and NaCl was added to a final concentration of 250 mM.

    Filtration:

    Article Title: Generation of Differentially Modified Microtubules Using in Vitro Enzymatic Approaches
    Article Snippet: .. Solutions required for α-TAT Purification 10X phosphate buffered saline (PBS). α-TAT resuspension buffer: 1X PBS supplemented with 10 mM MgCl2, 1 mM phenylmethylsulfonyl fluoride (PMSF), 5 mM dithiothreitol (DTT), protease inhibitor cocktail (Roche Applied Science). α-TAT GST-A buffer: 50 mM Tris-HCl (pH 7.5), 500 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT GST-B buffer: 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT ion exchange buffer A: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT. α-TAT ion exchange buffer B: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT, 2 M NaCl. α-TAT gel filtration buffer: 50 mM Tris-HCl (pH 7.5), 200 mM NaCl, 5 mM MgCl2, 2 mM tris(2-carboxyethyl)phosphine (TCEP). ..

    Protease Inhibitor:

    Article Title: PIE-1 promotes SUMOylation and activation of HDAC1 during the C. elegans oogenesis
    Article Snippet: .. ImmunoprecipitationEither synchronous adult worms (~200,000 animals) or early embryos (bleached from 10×100,000 adult animals) were collected and washed three times with M9 buffer before being homogenized in lysis buffer [20mM HEPES pH 7.5, 125mM Na3C6H5O7 (sodium citrate), 0.1%(v/v) Tween 20, 0.5%(v/v) Triton X-100, 2mM MgCl2, 1mM DTT, and a Mini Protease Inhibitor Cocktail Tablet (Roche)] using a FastPrep-24 benchtop homogenizer (MP Biomedicals). .. Worm or embryo lysates were centrifuged twice at 14,000×g for 30 min at 4°C, and supernatants were incubated with GFP-binding protein (GBP) beads for 1.5 h at 4°C on a rotating shaker.

    Article Title: A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors
    Article Snippet: .. Solutions and media SOC medium: 2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2 and 20 mM glucose SOB-ampicillin medium: 2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2 and 100 mg/ml ampicillin LB-ampicillin plate: 1% tryptone, 0.5% yeast extract, 1% NaCl, 10 mM MgCl2 , 100 mg/ml ampicillin and 1.3% agar Cell lysis solution: phosphate buffered saline (PBS), 0.5 mM DTT and a protease inhibitor cocktail (Roche) .. Protocol Transfer 1 ng of plasmid DNA in 25 ml of BL21 competent cell suspension.

    Article Title: The FA Core Complex Contains a Homo-dimeric Catalytic Module for the Symmetric Mono-ubiquitination of FANCI-FANCD2
    Article Snippet: .. Cells were collected post-infection by centrifugation (5,000 rpm, 4°C, 15 min), washed in PBS buffer supplemented with 5 mM MgCl2 , and resuspended in lysis buffer (25 mM HEPES NaOH [pH 7.6], 15 mM NaCl, 0.02% Tween 20, 10% glycerol, 1 mM EDTA, 1 mM EGTA, 2 mM MgCl2 , 2 mM 2-BME, 0.4 mM PMSF, and Roche protease inhibitor cocktail). .. The suspension was lysed by Dounce homogenization (40 strokes, tight pestle), and NaCl was added to a final concentration of 250 mM.

    Article Title: The common ancestral core of vertebrate and fungal telomerase RNAs
    Article Snippet: .. The nuclear pellet was washed once with wash buffer [50 mM Tris–HCl (pH 7.5), 5 mM MgCl2 , 10 mM β-mercaptoethanol, 20% glycerol and 0.6 M sucrose], resuspended and lysed in CHAPS lysis buffer [10 mM Tris–HCl (pH 7.5), 400 mM NaCl, 0.5% CHAPS, 5% glycerol, 1 mM ethylene glycol tetraacetic acid (EGTA), 1 mM MgCl2 , supplemented with 5 mM β-mercaptoethanol and 1× protease inhibitor cocktail (Roche)]. .. The nuclear extract was cleared by centrifugation at 14 000g for 10 min. Two milliliters of nuclear extract was immediately applied to an XK26/60 Sephacryl S-500 gel filtration column (GE Healthcare) equilibrated with column buffer [10 mM Tris–HCl (pH 7.5), 400 mM NaCl, 5% glycerol].

    Article Title: Generation of Differentially Modified Microtubules Using in Vitro Enzymatic Approaches
    Article Snippet: .. Solutions required for α-TAT Purification 10X phosphate buffered saline (PBS). α-TAT resuspension buffer: 1X PBS supplemented with 10 mM MgCl2, 1 mM phenylmethylsulfonyl fluoride (PMSF), 5 mM dithiothreitol (DTT), protease inhibitor cocktail (Roche Applied Science). α-TAT GST-A buffer: 50 mM Tris-HCl (pH 7.5), 500 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT GST-B buffer: 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT ion exchange buffer A: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT. α-TAT ion exchange buffer B: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT, 2 M NaCl. α-TAT gel filtration buffer: 50 mM Tris-HCl (pH 7.5), 200 mM NaCl, 5 mM MgCl2, 2 mM tris(2-carboxyethyl)phosphine (TCEP). ..

    Purification:

    Article Title: Generation of Differentially Modified Microtubules Using in Vitro Enzymatic Approaches
    Article Snippet: .. Solutions required for α-TAT Purification 10X phosphate buffered saline (PBS). α-TAT resuspension buffer: 1X PBS supplemented with 10 mM MgCl2, 1 mM phenylmethylsulfonyl fluoride (PMSF), 5 mM dithiothreitol (DTT), protease inhibitor cocktail (Roche Applied Science). α-TAT GST-A buffer: 50 mM Tris-HCl (pH 7.5), 500 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT GST-B buffer: 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 10 mM MgCl2, 5 mM DTT. α-TAT ion exchange buffer A: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT. α-TAT ion exchange buffer B: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 2 mM DTT, 2 M NaCl. α-TAT gel filtration buffer: 50 mM Tris-HCl (pH 7.5), 200 mM NaCl, 5 mM MgCl2, 2 mM tris(2-carboxyethyl)phosphine (TCEP). ..

    Incubation:

    Article Title: Identification of the BRD1 interaction network and its impact on mental disorder risk
    Article Snippet: .. Beads were added to the pre-cleared supernatant and incubated at 4 °C overnight before they were collected at 400 rpm for 15 min. Beads were washed twice in 10 mL Triton X-100 buffer 250 (20 mM Tris–HCl [pH 7.4], 250 mM NaCl, 10 mM MgCl2 , 2 mM EDTA, 10 % glycerol, 1 % Triton X-100, 2.5 mM β-glycerophosphate, 1 mM NaF, 1 mM DTT, protease inhibitors [Roche]) then moved to Protein LoBind tubes (Eppendorf, Hamburg, Germany) and then washed six times in Triton X-100 buffer 150 (20 mM Tris–HCl [pH 7.4], 150 mM NaCl, 10 mM MgCl2 , 2 mM EDTA, 10 % glycerol, 1 % Triton X-100, 2.5 mM β-glycerophosphate,1 mM NaF, 1 mM DTT, protease inhibitors [Roche]). .. Proteins were eluted in 100 μL Elution buffer (HENG) (10 mM HEPES-KOH [pH 9.0], 1.5 mM MgCl2, 0.25 mM EDTA, 20 % glycerol, 250 mM KCl, 0.3 % NP40 + 0.5 mg/mL V5 peptide [Sigma], protease inhibitors [Roche]).

    Lysis:

    Article Title: PIE-1 promotes SUMOylation and activation of HDAC1 during the C. elegans oogenesis
    Article Snippet: .. ImmunoprecipitationEither synchronous adult worms (~200,000 animals) or early embryos (bleached from 10×100,000 adult animals) were collected and washed three times with M9 buffer before being homogenized in lysis buffer [20mM HEPES pH 7.5, 125mM Na3C6H5O7 (sodium citrate), 0.1%(v/v) Tween 20, 0.5%(v/v) Triton X-100, 2mM MgCl2, 1mM DTT, and a Mini Protease Inhibitor Cocktail Tablet (Roche)] using a FastPrep-24 benchtop homogenizer (MP Biomedicals). .. Worm or embryo lysates were centrifuged twice at 14,000×g for 30 min at 4°C, and supernatants were incubated with GFP-binding protein (GBP) beads for 1.5 h at 4°C on a rotating shaker.

    Article Title: A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors
    Article Snippet: .. Solutions and media SOC medium: 2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2 and 20 mM glucose SOB-ampicillin medium: 2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2 and 100 mg/ml ampicillin LB-ampicillin plate: 1% tryptone, 0.5% yeast extract, 1% NaCl, 10 mM MgCl2 , 100 mg/ml ampicillin and 1.3% agar Cell lysis solution: phosphate buffered saline (PBS), 0.5 mM DTT and a protease inhibitor cocktail (Roche) .. Protocol Transfer 1 ng of plasmid DNA in 25 ml of BL21 competent cell suspension.

    Article Title: The FA Core Complex Contains a Homo-dimeric Catalytic Module for the Symmetric Mono-ubiquitination of FANCI-FANCD2
    Article Snippet: .. Cells were collected post-infection by centrifugation (5,000 rpm, 4°C, 15 min), washed in PBS buffer supplemented with 5 mM MgCl2 , and resuspended in lysis buffer (25 mM HEPES NaOH [pH 7.6], 15 mM NaCl, 0.02% Tween 20, 10% glycerol, 1 mM EDTA, 1 mM EGTA, 2 mM MgCl2 , 2 mM 2-BME, 0.4 mM PMSF, and Roche protease inhibitor cocktail). .. The suspension was lysed by Dounce homogenization (40 strokes, tight pestle), and NaCl was added to a final concentration of 250 mM.

    Article Title: The common ancestral core of vertebrate and fungal telomerase RNAs
    Article Snippet: .. The nuclear pellet was washed once with wash buffer [50 mM Tris–HCl (pH 7.5), 5 mM MgCl2 , 10 mM β-mercaptoethanol, 20% glycerol and 0.6 M sucrose], resuspended and lysed in CHAPS lysis buffer [10 mM Tris–HCl (pH 7.5), 400 mM NaCl, 0.5% CHAPS, 5% glycerol, 1 mM ethylene glycol tetraacetic acid (EGTA), 1 mM MgCl2 , supplemented with 5 mM β-mercaptoethanol and 1× protease inhibitor cocktail (Roche)]. .. The nuclear extract was cleared by centrifugation at 14 000g for 10 min. Two milliliters of nuclear extract was immediately applied to an XK26/60 Sephacryl S-500 gel filtration column (GE Healthcare) equilibrated with column buffer [10 mM Tris–HCl (pH 7.5), 400 mM NaCl, 5% glycerol].

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  • 92
    Roche tris buffer
    Localization of TcdE in E. coli and C. difficile . A. Cytoplasmic and membrane proteins analysis of E. coli lysogens of λCmrΔ(SR) carrying pBR322 (control) or pCD463 (+ tcdE -6xHis). B. Cytoplasmic and membrane proteins analysis of C. difficile strain carrying either pMTL84151 (control) or pRG46 (+ tcdE -6xHis). (1) SDS-PAGE coomassie stained gel. Western blots probed with 6XHis Tag antibody (2), ATPase Beta subunit antibody (3), and Ribosomal subunits LI/L2 monoclonal antibody (4). C. Membrane protein samples from bacterial cells expressing TcdE-6His <t>resuspended</t> in denature or native sample buffers and analyzed by Western blot using His-Tag antibody. D. Membrane proteins of JIR8094, TcdE mutant and complemented TcdE mutant strains were harvested from bacterial cultures induced with 20 ng/ml of ATc for 2 hours, separated in 16% <t>Tris-Glycine</t> gel and transferred into PVDF membrane. Panels 1. Ponceau stained membrane; 2. Probed with TcdE antibody.
    Tris Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 106 article reviews
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    92
    Roche immunoprecipitation buffer
    Vincristine induces resistance in RMS cells through BID inhibition by MCL-1. (A) Western blot results of the unbound fraction after MCL-1 <t>immunoprecipitation.</t> High efficiency of MCL-1 immunoprecipitation compared to Rabbit IgG control antibody. (B) Western blot results showing MCL-1 levels in CW9019 cell lysates (incubated with vincristine or DMSO for 36 hours) before performing the immunoprecipitation. (C) Left panel: Western blot results of the co-immunoprecipitation between MCL-1 and BID. Right panel: Quantification of the optical density of each protein and represented as binding ratio between BID and MCL-1. Results showed a significant increase in BID and MCL-1 binding after vincristine treatment, which is restored to control levels after the addition of S63845. Values indicate mean values ± SEM from at least three independent experiments, * p
    Immunoprecipitation Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 273 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation buffer/product/Roche
    Average 92 stars, based on 273 article reviews
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    92
    Roche atp level
    Effect of glucose and lactate on cellular <t>ATP</t> content in <t>IPEC-1</t> cells after 1 h treatment. IPEC-1 cells were grown to confluence (7 d) and treated for 1 h with HBSS-based solutions. Supernatant was removed and ATP content was measured by a luminescence based assay. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p
    Atp Level, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Roche or 2 calcium free medium
    Regulation of proteolytically cleaved ENaC by cpt-cAMP. A. Representative current trace from an oocyte preincubated with plasmin (10 μg/ml) for 3 hrs in <t>OR-2</t> medium. B. Average increase in both I max and I 30s levels. C . Current ratio. n=12.
    Or 2 Calcium Free Medium, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Localization of TcdE in E. coli and C. difficile . A. Cytoplasmic and membrane proteins analysis of E. coli lysogens of λCmrΔ(SR) carrying pBR322 (control) or pCD463 (+ tcdE -6xHis). B. Cytoplasmic and membrane proteins analysis of C. difficile strain carrying either pMTL84151 (control) or pRG46 (+ tcdE -6xHis). (1) SDS-PAGE coomassie stained gel. Western blots probed with 6XHis Tag antibody (2), ATPase Beta subunit antibody (3), and Ribosomal subunits LI/L2 monoclonal antibody (4). C. Membrane protein samples from bacterial cells expressing TcdE-6His resuspended in denature or native sample buffers and analyzed by Western blot using His-Tag antibody. D. Membrane proteins of JIR8094, TcdE mutant and complemented TcdE mutant strains were harvested from bacterial cultures induced with 20 ng/ml of ATc for 2 hours, separated in 16% Tris-Glycine gel and transferred into PVDF membrane. Panels 1. Ponceau stained membrane; 2. Probed with TcdE antibody.

    Journal: PLoS Pathogens

    Article Title: Secretion of Clostridium difficile Toxins A and B Requires the Holin-like Protein TcdE

    doi: 10.1371/journal.ppat.1002727

    Figure Lengend Snippet: Localization of TcdE in E. coli and C. difficile . A. Cytoplasmic and membrane proteins analysis of E. coli lysogens of λCmrΔ(SR) carrying pBR322 (control) or pCD463 (+ tcdE -6xHis). B. Cytoplasmic and membrane proteins analysis of C. difficile strain carrying either pMTL84151 (control) or pRG46 (+ tcdE -6xHis). (1) SDS-PAGE coomassie stained gel. Western blots probed with 6XHis Tag antibody (2), ATPase Beta subunit antibody (3), and Ribosomal subunits LI/L2 monoclonal antibody (4). C. Membrane protein samples from bacterial cells expressing TcdE-6His resuspended in denature or native sample buffers and analyzed by Western blot using His-Tag antibody. D. Membrane proteins of JIR8094, TcdE mutant and complemented TcdE mutant strains were harvested from bacterial cultures induced with 20 ng/ml of ATc for 2 hours, separated in 16% Tris-Glycine gel and transferred into PVDF membrane. Panels 1. Ponceau stained membrane; 2. Probed with TcdE antibody.

    Article Snippet: C. difficile toxin and Lactate dehydrogenase (LDH) assays Culture supernatants were collected and filtered, and the cell pellets were resuspended in 10 mM Tris buffer, pH 8.0 containing a protease inhibitor cocktail (Roche, Mannheim, Germany).

    Techniques: SDS Page, Staining, Western Blot, Expressing, Mutagenesis

    Vincristine induces resistance in RMS cells through BID inhibition by MCL-1. (A) Western blot results of the unbound fraction after MCL-1 immunoprecipitation. High efficiency of MCL-1 immunoprecipitation compared to Rabbit IgG control antibody. (B) Western blot results showing MCL-1 levels in CW9019 cell lysates (incubated with vincristine or DMSO for 36 hours) before performing the immunoprecipitation. (C) Left panel: Western blot results of the co-immunoprecipitation between MCL-1 and BID. Right panel: Quantification of the optical density of each protein and represented as binding ratio between BID and MCL-1. Results showed a significant increase in BID and MCL-1 binding after vincristine treatment, which is restored to control levels after the addition of S63845. Values indicate mean values ± SEM from at least three independent experiments, * p

    Journal: bioRxiv

    Article Title: Sequential combinations of chemotherapeutic agents with BH3 mimetics to treat rhabdomyosarcoma and avoid resistance

    doi: 10.1101/2020.01.24.918532

    Figure Lengend Snippet: Vincristine induces resistance in RMS cells through BID inhibition by MCL-1. (A) Western blot results of the unbound fraction after MCL-1 immunoprecipitation. High efficiency of MCL-1 immunoprecipitation compared to Rabbit IgG control antibody. (B) Western blot results showing MCL-1 levels in CW9019 cell lysates (incubated with vincristine or DMSO for 36 hours) before performing the immunoprecipitation. (C) Left panel: Western blot results of the co-immunoprecipitation between MCL-1 and BID. Right panel: Quantification of the optical density of each protein and represented as binding ratio between BID and MCL-1. Results showed a significant increase in BID and MCL-1 binding after vincristine treatment, which is restored to control levels after the addition of S63845. Values indicate mean values ± SEM from at least three independent experiments, * p

    Article Snippet: Immunoprecipitation Cells were lysed in Immunoprecipitation buffer (150 mM NaCl, 10 mM Hepes, 2 mM EDTA, 1% Triton, 1.5 mM MgCl2, 10% glycerol, protease inhibitor from Roche) and centrifuged at 14,000 × g , 15 minutes at 4°C.

    Techniques: Inhibition, Western Blot, Immunoprecipitation, Incubation, Binding Assay

    Effect of glucose and lactate on cellular ATP content in IPEC-1 cells after 1 h treatment. IPEC-1 cells were grown to confluence (7 d) and treated for 1 h with HBSS-based solutions. Supernatant was removed and ATP content was measured by a luminescence based assay. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p

    Journal: PLoS ONE

    Article Title: Physiological Concentration of Exogenous Lactate Reduces Antimycin A Triggered Oxidative Stress in Intestinal Epithelial Cell Line IPEC-1 and IPEC-J2 In Vitro

    doi: 10.1371/journal.pone.0153135

    Figure Lengend Snippet: Effect of glucose and lactate on cellular ATP content in IPEC-1 cells after 1 h treatment. IPEC-1 cells were grown to confluence (7 d) and treated for 1 h with HBSS-based solutions. Supernatant was removed and ATP content was measured by a luminescence based assay. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p

    Article Snippet: Cellular ATP Level in IPEC-1 Cells The effect of lactate on cellular ATP level was quantified using a commercial ATP kit (ATP Bioluminescence Assay Kit HS II, Roche).

    Techniques: Luminescence Assay

    Regulation of proteolytically cleaved ENaC by cpt-cAMP. A. Representative current trace from an oocyte preincubated with plasmin (10 μg/ml) for 3 hrs in OR-2 medium. B. Average increase in both I max and I 30s levels. C . Current ratio. n=12.

    Journal: Biochimica et biophysica acta

    Article Title: Cpt-cAMP activates human epithelial sodium channels via relieving self-inhibition

    doi: 10.1016/j.bbamem.2011.03.004

    Figure Lengend Snippet: Regulation of proteolytically cleaved ENaC by cpt-cAMP. A. Representative current trace from an oocyte preincubated with plasmin (10 μg/ml) for 3 hrs in OR-2 medium. B. Average increase in both I max and I 30s levels. C . Current ratio. n=12.

    Article Snippet: Ovary lobes were removed and digested in OR-2 calcium-free medium (in mM: 82.5 NaCl, 2.5 KCl, 1.0 MgCl2 , 1.0 Na2 HPO4 , and 10.0 HEPES, pH 7.5) with the addition of 2 mg/ml collagenase (Roche Indianapolis).

    Techniques: Cycling Probe Technology