phosphatase inhibitors  (Roche)


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    Structured Review

    Roche phosphatase inhibitors
    Phosphatase Inhibitors, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 638 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphatase inhibitors/product/Roche
    Average 94 stars, based on 638 article reviews
    Price from $9.99 to $1999.99
    phosphatase inhibitors - by Bioz Stars, 2020-09
    94/100 stars

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    Related Articles

    Co-Immunoprecipitation Assay:

    Article Title: Distinct and sequential re-replication barriers ensure precise genome duplication
    Article Snippet: .. For co-immunoprecipitation assays, cells were lysed in Co-IP buffer (50 mM HEPES pH 7.2, 33 mM KAc, 1 mM MgCl2 , 0.5% triton X-100, and 10% glycerol) containing protease inhibitors (0.1 mM AEBSF, 10 µg/mL pepstatin A, 10 µg/mL leupeptin, 10 µg/mL aprotinin), phosphatase inhibitors (5 µg/mL phosvitin, 1 mM β-glycerol phosphate, 1 mM Na-orthovanadate), 1 mM ATP, and supplemented with 5 mM CaCl2 and 15 units of S7 micrococcal nuclease (Roche). .. The supernatants were incubated and rotated with Protein A beads (Roche) with an anti-Mcm2 antibody (Bethyl, 1:1000) at 4°C with rotation for 4 hours.

    Radio Immunoprecipitation:

    Article Title: Long Noncoding RNA LINC00460 Promotes the Gefitinib Resistance of Nonsmall Cell Lung Cancer Through Epidermal Growth Factor Receptor by Sponging miR-769-5p
    Article Snippet: .. Western blotting Cells were treated with radio-immunoprecipitation assay lysis reagent (Millipore, Billerica, MA), and the lysates were prepared with phosphatase inhibitors and protease inhibitors (Roche, Basel, Switzerland). .. BCA Protein Assay Kit (Thermo Fisher) was used to determine the protein concentration.

    Protease Inhibitor:

    Article Title: The miR-181d-regulated metalloproteinase Adamts1 enzymatically impairs adipogenesis via ECM remodeling
    Article Snippet: .. Adipose tissue and cultured cells were scraped into lysis buffer containing 50 mM Tris-HCl (pH 6.8), 2% SDS, phosphatase inhibitors (10 mM Na3 VO4 , 10 mM NaF), and a protease inhibitor cocktail (Roche, Mannheim, Germany). .. Equal amounts of protein were subjected to SDS/PAGE and immunoblotted with specific primary antibodies against one of the following targets: Adamts1 (Millipore, Billerica, MA, USA); Hsp90, PPAR γ , or C/EBP α , (Santa Cruz, CA, USA); 422/aP2 (prepared in Dr. Lane's lab); p38 MAPK kinase, phospho-p38, Smad1, phospho-Smad1/5/8, FAK, phospho-FAK, ERK1/2, and phospho-ERK1/2(Cell Signaling Technology, Danvers, MA, USA); Pref1 (Proteintech Group, Chicago, IL, USA); Col5 α 1 (ABclonal, Cambridge, MA, USA); Thbs1 (Novus, Littleton, CO, USA); Fibronectin (BD, Franklin Lakes, NJ, USA); Col1 α 1 and β -actin (Sigma-Aldrich).

    Article Title: Erythropoietin-driven signalling and cell migration mediated by polyADP-ribosylation
    Article Snippet: .. Cells (2 × 106 ) were lysed at 4 °C in lysis buffer (50 mℳ Tris pH 7.4, 1% Triton X-100, 5 mℳ EDTA, 150 mℳ NaCl) containing phosphatase inhibitors (1 mℳ vanadate, 50 mℳ sodium fluoride, 10 mℳ sodium pyrophosphate and 1 mℳ ZnCl2 , final concentrations) and protease inhibitors (Complete Protease Inhibitor Cocktail, Roche Diagnostics, Basel, Switzerland). .. For nuclear extraction, cells (6 × 106 ) were lysed at 4 °C in lysis buffer (10 mℳ Tris–HCl pH 7.5, 10 mℳ KCl, 0.1 mℳ EDTA, 1.5 mℳ MgCl2 , 0.2% NP-40, 1 mℳ DTT) containing phosphatase and protease inhibitors (as above).

    Article Title: Growing Rats Respond to a Sulfur Amino Acid-Deficient Diet by Phosphorylation of the ? Subunit of Eukaryotic Initiation Factor 2 Heterotrimeric Complex and Induction of Adaptive Components of the Integrated Stress Response 1Growing Rats Respond to a Sulfur Amino Acid-Deficient Diet by Phosphorylation of the ? Subunit of Eukaryotic Initiation Factor 2 Heterotrimeric Complex and Induction of Adapti
    Article Snippet: .. Rat liver was homogenized in TNESV lysis buffer supplemented with protease and phosphatase inhibitors [50 mmol/L Tris, pH 7.5, 1% (v:v) Nonidet P-40, 2 mmol/L EDTA, 150 mmol/L NaCl, and 10 mmol/L sodium orthovanadate] supplemented with 1× PhosSTOP phosphatase inhibitor cocktail (Roche Applied Science) and 1× Complete Protease Inhibitor Cocktail (Roche), to form 20% (wt:v) homogenates. ..

    Cell Culture:

    Article Title: The miR-181d-regulated metalloproteinase Adamts1 enzymatically impairs adipogenesis via ECM remodeling
    Article Snippet: .. Adipose tissue and cultured cells were scraped into lysis buffer containing 50 mM Tris-HCl (pH 6.8), 2% SDS, phosphatase inhibitors (10 mM Na3 VO4 , 10 mM NaF), and a protease inhibitor cocktail (Roche, Mannheim, Germany). .. Equal amounts of protein were subjected to SDS/PAGE and immunoblotted with specific primary antibodies against one of the following targets: Adamts1 (Millipore, Billerica, MA, USA); Hsp90, PPAR γ , or C/EBP α , (Santa Cruz, CA, USA); 422/aP2 (prepared in Dr. Lane's lab); p38 MAPK kinase, phospho-p38, Smad1, phospho-Smad1/5/8, FAK, phospho-FAK, ERK1/2, and phospho-ERK1/2(Cell Signaling Technology, Danvers, MA, USA); Pref1 (Proteintech Group, Chicago, IL, USA); Col5 α 1 (ABclonal, Cambridge, MA, USA); Thbs1 (Novus, Littleton, CO, USA); Fibronectin (BD, Franklin Lakes, NJ, USA); Col1 α 1 and β -actin (Sigma-Aldrich).

    Western Blot:

    Article Title: Long Noncoding RNA LINC00460 Promotes the Gefitinib Resistance of Nonsmall Cell Lung Cancer Through Epidermal Growth Factor Receptor by Sponging miR-769-5p
    Article Snippet: .. Western blotting Cells were treated with radio-immunoprecipitation assay lysis reagent (Millipore, Billerica, MA), and the lysates were prepared with phosphatase inhibitors and protease inhibitors (Roche, Basel, Switzerland). .. BCA Protein Assay Kit (Thermo Fisher) was used to determine the protein concentration.

    Lysis:

    Article Title: The miR-181d-regulated metalloproteinase Adamts1 enzymatically impairs adipogenesis via ECM remodeling
    Article Snippet: .. Adipose tissue and cultured cells were scraped into lysis buffer containing 50 mM Tris-HCl (pH 6.8), 2% SDS, phosphatase inhibitors (10 mM Na3 VO4 , 10 mM NaF), and a protease inhibitor cocktail (Roche, Mannheim, Germany). .. Equal amounts of protein were subjected to SDS/PAGE and immunoblotted with specific primary antibodies against one of the following targets: Adamts1 (Millipore, Billerica, MA, USA); Hsp90, PPAR γ , or C/EBP α , (Santa Cruz, CA, USA); 422/aP2 (prepared in Dr. Lane's lab); p38 MAPK kinase, phospho-p38, Smad1, phospho-Smad1/5/8, FAK, phospho-FAK, ERK1/2, and phospho-ERK1/2(Cell Signaling Technology, Danvers, MA, USA); Pref1 (Proteintech Group, Chicago, IL, USA); Col5 α 1 (ABclonal, Cambridge, MA, USA); Thbs1 (Novus, Littleton, CO, USA); Fibronectin (BD, Franklin Lakes, NJ, USA); Col1 α 1 and β -actin (Sigma-Aldrich).

    Article Title: Erythropoietin-driven signalling and cell migration mediated by polyADP-ribosylation
    Article Snippet: .. Cells (2 × 106 ) were lysed at 4 °C in lysis buffer (50 mℳ Tris pH 7.4, 1% Triton X-100, 5 mℳ EDTA, 150 mℳ NaCl) containing phosphatase inhibitors (1 mℳ vanadate, 50 mℳ sodium fluoride, 10 mℳ sodium pyrophosphate and 1 mℳ ZnCl2 , final concentrations) and protease inhibitors (Complete Protease Inhibitor Cocktail, Roche Diagnostics, Basel, Switzerland). .. For nuclear extraction, cells (6 × 106 ) were lysed at 4 °C in lysis buffer (10 mℳ Tris–HCl pH 7.5, 10 mℳ KCl, 0.1 mℳ EDTA, 1.5 mℳ MgCl2 , 0.2% NP-40, 1 mℳ DTT) containing phosphatase and protease inhibitors (as above).

    Article Title: Growing Rats Respond to a Sulfur Amino Acid-Deficient Diet by Phosphorylation of the ? Subunit of Eukaryotic Initiation Factor 2 Heterotrimeric Complex and Induction of Adaptive Components of the Integrated Stress Response 1Growing Rats Respond to a Sulfur Amino Acid-Deficient Diet by Phosphorylation of the ? Subunit of Eukaryotic Initiation Factor 2 Heterotrimeric Complex and Induction of Adapti
    Article Snippet: .. Rat liver was homogenized in TNESV lysis buffer supplemented with protease and phosphatase inhibitors [50 mmol/L Tris, pH 7.5, 1% (v:v) Nonidet P-40, 2 mmol/L EDTA, 150 mmol/L NaCl, and 10 mmol/L sodium orthovanadate] supplemented with 1× PhosSTOP phosphatase inhibitor cocktail (Roche Applied Science) and 1× Complete Protease Inhibitor Cocktail (Roche), to form 20% (wt:v) homogenates. ..

    Article Title: Long Noncoding RNA LINC00460 Promotes the Gefitinib Resistance of Nonsmall Cell Lung Cancer Through Epidermal Growth Factor Receptor by Sponging miR-769-5p
    Article Snippet: .. Western blotting Cells were treated with radio-immunoprecipitation assay lysis reagent (Millipore, Billerica, MA), and the lysates were prepared with phosphatase inhibitors and protease inhibitors (Roche, Basel, Switzerland). .. BCA Protein Assay Kit (Thermo Fisher) was used to determine the protein concentration.

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  • 86
    Roche lambda phosphatase assay
    S6K1 phosphorylate FMRP S499 in vitro . ( A ) The specificity of the C-terminal, phospho-insensitive FMRP tFMRP antibody (Abcam #17722) was verified by immunoblotting whole cell cortical lysates from 2 month-old Fmr1 y/+ (WT) and Fmr1 y/− (KO) mice. Short and long exposures (exp.) are shown. Arrows point to three FMRP-specific bands while asterisks point to nonspecific bands. Molecular weight is indicated by the colored ladders on the left which refer to panels A–D ( B ) Fmr1 y/+ and Fmr1 y/− cortical lysates were immunoblotted with a pFMRP antibody and the film exposed for 1 minute (PhosphoSolution #p1125-499). The membrane was then stripped and reprobed with secondary only antibody (1 minute exposure) to verify removal of the primary antibody. The membrane was then probed with tFMRP antibody (30 second exposure). The arrow points to the FMRP-specific band while asterisks point to two nonspecific pFMRP bands. ( C ) Cortical lysate from Fmr1 y/+ mice was incubated with or without <t>lambda</t> (λ) phosphatase for 30 minutes and immunoblotted with pFMRP, stripped and reprobed with tFMRP. ( D ) Neuro2a cell lysate (N2a) and unphosphorylated recombinant human FMRP (rFMRP) were immunoblotted with pFMRP. ( E ) GST-FMRP S499, S499A and S499D were transfected into N2a cells and lysates analyzed by immunoblot 24 hours later. tFMRP recognized all three isoforms (black, GST-labeled arrow) as well as endogenous FMRP (white arrows) however pFMRP only recognized S499 and endogenous FMRP from the same membrane. ( F ) Recombinant FMRP was incubated with ATP and with, or without (Neg), S6K1 or S6K2 and immunoblotted for pFMRP or total FMRP (tFMRP = 1C3 antibody here). The experiments were reproduced in triplicate.
    Lambda Phosphatase Assay, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Roche sds lysis buffer
    LRRK2 forms kinase-sensitive high molecular weight conformations. A , cell pellets derived from four lymphoblast cell lines (labeled Lymph1 to -4 ) were split equally for lysis by freeze/thaw cycles directly in <t>PBS</t> or lysis with 1% <t>SDS</t> and PBS with sonication,
    Sds Lysis Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche phosstop phosphatase inhibitor cocktail
    The stability of the CjWRKY1 protein in C. japonica 156-S protoplasts. ( a ) The different stabilities of WT and mutant CjWRKY1 proteins. CjWRKY1 proteins were detected using CjWRKY1-specific antibodies. ( b ) A proteasome inhibitor, MG132 (50 μM), stabilized the CjWRKY1 proteins for 48 h. ( c ) Synergistic effects of MG132, protease inhibitors, and protein phosphatase inhibitors on the accumulation of CjWRKY1 proteins. Cj156-S protoplasts were treated with 50 μM MG132, 1 mg/ml complete EDTA-free protease inhibitor cocktail, and/or 5 mg/ml <t>phosSTOP</t> phosphatase inhibitor cocktail. An acrylamide gel containing 50 μM Phos-tag was used to detect phosphorylated WRKY1. Arrows indicate the shifted bands corresponding to phosphorylated CjWRKY1. DMSO (0.1%) was used for mock treatments in ( b , c ). All experiments were repeated at least three times with similar results.
    Phosstop Phosphatase Inhibitor Cocktail, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 242 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosstop phosphatase inhibitor cocktail/product/Roche
    Average 94 stars, based on 242 article reviews
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    86
    Roche calcineurin activity anesthetized mice
    Depletion of Calstabin2 causes intracellular Ca 2+ leakage, activation of <t>calcineurin</t> and AKT-mTOR pathway. (A), Resting Ca 2+ determined by the ratio of F340/F380 fluorescence in WT and KO mice at different ages. At 48 weeks, resting [Ca 2+ ] i was 20% higher in KO cells than in WT controls. Numbers in the bars indicate the number of the analyzed cells isolated from five to six mice. (B), Calcineurin activity was 48% higher in aged KO mice than in the age-matched WT mice and 1.8-fold higher than in young KO mice. Immunoblots for proteins involved in AKT-mTOR signaling pathway in hearts from 12-week-old (C) and 48-week-old (D) mice. The graphs indicate the relative expression levels of p-AKT, p-p70S6K and p-mTOR. n = 5 per group. Quantitative data are shown as means ± SEM. *P
    Calcineurin Activity Anesthetized Mice, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    S6K1 phosphorylate FMRP S499 in vitro . ( A ) The specificity of the C-terminal, phospho-insensitive FMRP tFMRP antibody (Abcam #17722) was verified by immunoblotting whole cell cortical lysates from 2 month-old Fmr1 y/+ (WT) and Fmr1 y/− (KO) mice. Short and long exposures (exp.) are shown. Arrows point to three FMRP-specific bands while asterisks point to nonspecific bands. Molecular weight is indicated by the colored ladders on the left which refer to panels A–D ( B ) Fmr1 y/+ and Fmr1 y/− cortical lysates were immunoblotted with a pFMRP antibody and the film exposed for 1 minute (PhosphoSolution #p1125-499). The membrane was then stripped and reprobed with secondary only antibody (1 minute exposure) to verify removal of the primary antibody. The membrane was then probed with tFMRP antibody (30 second exposure). The arrow points to the FMRP-specific band while asterisks point to two nonspecific pFMRP bands. ( C ) Cortical lysate from Fmr1 y/+ mice was incubated with or without lambda (λ) phosphatase for 30 minutes and immunoblotted with pFMRP, stripped and reprobed with tFMRP. ( D ) Neuro2a cell lysate (N2a) and unphosphorylated recombinant human FMRP (rFMRP) were immunoblotted with pFMRP. ( E ) GST-FMRP S499, S499A and S499D were transfected into N2a cells and lysates analyzed by immunoblot 24 hours later. tFMRP recognized all three isoforms (black, GST-labeled arrow) as well as endogenous FMRP (white arrows) however pFMRP only recognized S499 and endogenous FMRP from the same membrane. ( F ) Recombinant FMRP was incubated with ATP and with, or without (Neg), S6K1 or S6K2 and immunoblotted for pFMRP or total FMRP (tFMRP = 1C3 antibody here). The experiments were reproduced in triplicate.

    Journal: PLoS ONE

    Article Title: FMRP S499 Is Phosphorylated Independent of mTORC1-S6K1 Activity

    doi: 10.1371/journal.pone.0096956

    Figure Lengend Snippet: S6K1 phosphorylate FMRP S499 in vitro . ( A ) The specificity of the C-terminal, phospho-insensitive FMRP tFMRP antibody (Abcam #17722) was verified by immunoblotting whole cell cortical lysates from 2 month-old Fmr1 y/+ (WT) and Fmr1 y/− (KO) mice. Short and long exposures (exp.) are shown. Arrows point to three FMRP-specific bands while asterisks point to nonspecific bands. Molecular weight is indicated by the colored ladders on the left which refer to panels A–D ( B ) Fmr1 y/+ and Fmr1 y/− cortical lysates were immunoblotted with a pFMRP antibody and the film exposed for 1 minute (PhosphoSolution #p1125-499). The membrane was then stripped and reprobed with secondary only antibody (1 minute exposure) to verify removal of the primary antibody. The membrane was then probed with tFMRP antibody (30 second exposure). The arrow points to the FMRP-specific band while asterisks point to two nonspecific pFMRP bands. ( C ) Cortical lysate from Fmr1 y/+ mice was incubated with or without lambda (λ) phosphatase for 30 minutes and immunoblotted with pFMRP, stripped and reprobed with tFMRP. ( D ) Neuro2a cell lysate (N2a) and unphosphorylated recombinant human FMRP (rFMRP) were immunoblotted with pFMRP. ( E ) GST-FMRP S499, S499A and S499D were transfected into N2a cells and lysates analyzed by immunoblot 24 hours later. tFMRP recognized all three isoforms (black, GST-labeled arrow) as well as endogenous FMRP (white arrows) however pFMRP only recognized S499 and endogenous FMRP from the same membrane. ( F ) Recombinant FMRP was incubated with ATP and with, or without (Neg), S6K1 or S6K2 and immunoblotted for pFMRP or total FMRP (tFMRP = 1C3 antibody here). The experiments were reproduced in triplicate.

    Article Snippet: Lambda Phosphatase Assay A single CD1 adult mouse hippocampus was homogenized in 300 µl phosphatase assay lysis buffer (4 mM HEPES, 0.5% Triton-X-100, 120 mM NaCl, and 2 Roche protease inhibitor tablets per 10 ml).

    Techniques: In Vitro, Mouse Assay, Molecular Weight, Incubation, Recombinant, Transfection, Labeling

    LRRK2 forms kinase-sensitive high molecular weight conformations. A , cell pellets derived from four lymphoblast cell lines (labeled Lymph1 to -4 ) were split equally for lysis by freeze/thaw cycles directly in PBS or lysis with 1% SDS and PBS with sonication,

    Journal: The Journal of Biological Chemistry

    Article Title: Dependence of Leucine-rich Repeat Kinase 2 (LRRK2) Kinase Activity on Dimerization *

    doi: 10.1074/jbc.M109.025437

    Figure Lengend Snippet: LRRK2 forms kinase-sensitive high molecular weight conformations. A , cell pellets derived from four lymphoblast cell lines (labeled Lymph1 to -4 ) were split equally for lysis by freeze/thaw cycles directly in PBS or lysis with 1% SDS and PBS with sonication,

    Article Snippet: Cell pellets were resuspended with either native lysis buffer (phosphate-buffered saline (PBS), pH 7.4, 1× protease and phosphatase inhibitors (Roche Applied Science)), lysed by four cycles of freezing and thawing, and analyzed on a 3–12% bis-tris Blue native polyacrylamide gel (Invitrogen) or lysates combined with SDS-lysis buffer (PBS, pH 7.4, 1% SDS, 1× protease and phosphatase inhibitors (Roche Applied Science)) and sonicated with 10% power for 10 s on a Branson dismembranator and analyzed on a 7% Tris acetate SDS gel (Invitrogen).

    Techniques: Molecular Weight, Derivative Assay, Labeling, Lysis, Sonication

    The stability of the CjWRKY1 protein in C. japonica 156-S protoplasts. ( a ) The different stabilities of WT and mutant CjWRKY1 proteins. CjWRKY1 proteins were detected using CjWRKY1-specific antibodies. ( b ) A proteasome inhibitor, MG132 (50 μM), stabilized the CjWRKY1 proteins for 48 h. ( c ) Synergistic effects of MG132, protease inhibitors, and protein phosphatase inhibitors on the accumulation of CjWRKY1 proteins. Cj156-S protoplasts were treated with 50 μM MG132, 1 mg/ml complete EDTA-free protease inhibitor cocktail, and/or 5 mg/ml phosSTOP phosphatase inhibitor cocktail. An acrylamide gel containing 50 μM Phos-tag was used to detect phosphorylated WRKY1. Arrows indicate the shifted bands corresponding to phosphorylated CjWRKY1. DMSO (0.1%) was used for mock treatments in ( b , c ). All experiments were repeated at least three times with similar results.

    Journal: Scientific Reports

    Article Title: Tyrosine phosphorylation and protein degradation control the transcriptional activity of WRKY involved in benzylisoquinoline alkaloid biosynthesis

    doi: 10.1038/srep31988

    Figure Lengend Snippet: The stability of the CjWRKY1 protein in C. japonica 156-S protoplasts. ( a ) The different stabilities of WT and mutant CjWRKY1 proteins. CjWRKY1 proteins were detected using CjWRKY1-specific antibodies. ( b ) A proteasome inhibitor, MG132 (50 μM), stabilized the CjWRKY1 proteins for 48 h. ( c ) Synergistic effects of MG132, protease inhibitors, and protein phosphatase inhibitors on the accumulation of CjWRKY1 proteins. Cj156-S protoplasts were treated with 50 μM MG132, 1 mg/ml complete EDTA-free protease inhibitor cocktail, and/or 5 mg/ml phosSTOP phosphatase inhibitor cocktail. An acrylamide gel containing 50 μM Phos-tag was used to detect phosphorylated WRKY1. Arrows indicate the shifted bands corresponding to phosphorylated CjWRKY1. DMSO (0.1%) was used for mock treatments in ( b , c ). All experiments were repeated at least three times with similar results.

    Article Snippet: An equal volume of extraction buffer [50 mM Tris-HCl (pH 7.5), 1% SDS, 10% glycerol, 2 mM DTT, 1 mM EDTA, 50 μM MG132 (Millipore), complete EDTA-free protease inhibitor cocktail (Roche) and phosSTOP phosphatase inhibitor cocktail (Roche)] to the cell weight was added to powdered samples and placed on ice for 10 min. After centrifugation at 15,000 × g at 4 °C for 20 min, supernatants were recovered and used as total protein fractions.

    Techniques: Mutagenesis, Protease Inhibitor, Acrylamide Gel Assay

    Depletion of Calstabin2 causes intracellular Ca 2+ leakage, activation of calcineurin and AKT-mTOR pathway. (A), Resting Ca 2+ determined by the ratio of F340/F380 fluorescence in WT and KO mice at different ages. At 48 weeks, resting [Ca 2+ ] i was 20% higher in KO cells than in WT controls. Numbers in the bars indicate the number of the analyzed cells isolated from five to six mice. (B), Calcineurin activity was 48% higher in aged KO mice than in the age-matched WT mice and 1.8-fold higher than in young KO mice. Immunoblots for proteins involved in AKT-mTOR signaling pathway in hearts from 12-week-old (C) and 48-week-old (D) mice. The graphs indicate the relative expression levels of p-AKT, p-p70S6K and p-mTOR. n = 5 per group. Quantitative data are shown as means ± SEM. *P

    Journal: Scientific Reports

    Article Title: Functional Role of Calstabin2 in Age-related Cardiac Alterations

    doi: 10.1038/srep07425

    Figure Lengend Snippet: Depletion of Calstabin2 causes intracellular Ca 2+ leakage, activation of calcineurin and AKT-mTOR pathway. (A), Resting Ca 2+ determined by the ratio of F340/F380 fluorescence in WT and KO mice at different ages. At 48 weeks, resting [Ca 2+ ] i was 20% higher in KO cells than in WT controls. Numbers in the bars indicate the number of the analyzed cells isolated from five to six mice. (B), Calcineurin activity was 48% higher in aged KO mice than in the age-matched WT mice and 1.8-fold higher than in young KO mice. Immunoblots for proteins involved in AKT-mTOR signaling pathway in hearts from 12-week-old (C) and 48-week-old (D) mice. The graphs indicate the relative expression levels of p-AKT, p-p70S6K and p-mTOR. n = 5 per group. Quantitative data are shown as means ± SEM. *P

    Article Snippet: Immunoblotting and calcineurin activity Anesthetized mice were sacrificed immediately and mouse ventricles were harvested and homogenized in RIPA lysis buffer containing a protease inhibitor cocktail (Roche, Basel, Switzerland), proteins were resolved by SDS–PAGE and transferred to PVDF membranes (Millipore, Billerica, MA).

    Techniques: Activation Assay, Fluorescence, Mouse Assay, Isolation, Activity Assay, Western Blot, Expressing