Structured Review

Millipore m na3 vo4
M Na3 Vo4, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/m na3 vo4/product/Millipore
Average 99 stars, based on 10 article reviews
Price from $9.99 to $1999.99
m na3 vo4 - by Bioz Stars, 2020-09
99/100 stars

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Electrophoresis:

Article Title: A Novel Pathway for Mammary Epithelial Cell Invasion Induced by the Helix-Loop-Helix Protein Id-1
Article Snippet: .. Concentrated conditioned medium was mixed with nonreducing Laemmli sample buffer and incubated at 37°C for 15 min. After electrophoresis, the gels were incubated for 1 h in 2.5% Triton X-100 at room temperature, followed by 24 to 48 h in substrate buffer (100 mM Tris-HCl [pH 7.4]–15 mM CaCl2 ) in the absence or presence of GM6001 (0.2 mM in DMSO; supplied by Glycomed Corporation and obtained from Z. Werb [ ]), EDTA (10 mM), ortho -phenanthroline (1 mM in DMSO; Sigma), PMSF (5 mM), or AEBSF (0.5 mM; Calbiochem). .. The gels were stained with Coomassie blue for 30 min and were destained with 30% methanol–10% acetic acid.

Protease Inhibitor:

Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells
Article Snippet: .. Intracellular inhibition of protease digestion In order to inhibit intracellular protease activity, 5 × 106 cells/sample were treated with: Complete™ protease inhibitor tablets; either 1× (1 tablet for every 50 mL of media) or 2× (2 tablets for every 50 mL of media); or treated with antipain (2.0 μg/mL) plus leupeptin (2.0 μg/mL) (both from Sigma-Aldrich, St Louis, MO, USA) for cysteine protease inhibition; or aprotinin (2.0 μg/mL) plus PMSF (100 μg/mL) (both from Sigma-Aldrich) for serine protease inhibition; for to 24 hrs. ..

Incubation:

Article Title: Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past
Article Snippet: .. Supernatant samples of Bacillus sp. JR3 were also incubated in the presence of 1 mM PMSF (phenylmethylsulfonyl fluoride, Sigma) to measure the effect of this serine inhibitor on enzyme activity. .. Kinetic parameters (Vmax and Km) were determined under optimal assay conditions by fitting hyperbolic Michaelis-Menten curves with GraphPad Prism ® software version 6.

Article Title: A Novel Pathway for Mammary Epithelial Cell Invasion Induced by the Helix-Loop-Helix Protein Id-1
Article Snippet: .. Concentrated conditioned medium was mixed with nonreducing Laemmli sample buffer and incubated at 37°C for 15 min. After electrophoresis, the gels were incubated for 1 h in 2.5% Triton X-100 at room temperature, followed by 24 to 48 h in substrate buffer (100 mM Tris-HCl [pH 7.4]–15 mM CaCl2 ) in the absence or presence of GM6001 (0.2 mM in DMSO; supplied by Glycomed Corporation and obtained from Z. Werb [ ]), EDTA (10 mM), ortho -phenanthroline (1 mM in DMSO; Sigma), PMSF (5 mM), or AEBSF (0.5 mM; Calbiochem). .. The gels were stained with Coomassie blue for 30 min and were destained with 30% methanol–10% acetic acid.

Inhibition:

Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells
Article Snippet: .. Intracellular inhibition of protease digestion In order to inhibit intracellular protease activity, 5 × 106 cells/sample were treated with: Complete™ protease inhibitor tablets; either 1× (1 tablet for every 50 mL of media) or 2× (2 tablets for every 50 mL of media); or treated with antipain (2.0 μg/mL) plus leupeptin (2.0 μg/mL) (both from Sigma-Aldrich, St Louis, MO, USA) for cysteine protease inhibition; or aprotinin (2.0 μg/mL) plus PMSF (100 μg/mL) (both from Sigma-Aldrich) for serine protease inhibition; for to 24 hrs. ..

Activity Assay:

Article Title: Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past
Article Snippet: .. Supernatant samples of Bacillus sp. JR3 were also incubated in the presence of 1 mM PMSF (phenylmethylsulfonyl fluoride, Sigma) to measure the effect of this serine inhibitor on enzyme activity. .. Kinetic parameters (Vmax and Km) were determined under optimal assay conditions by fitting hyperbolic Michaelis-Menten curves with GraphPad Prism ® software version 6.

Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells
Article Snippet: .. Intracellular inhibition of protease digestion In order to inhibit intracellular protease activity, 5 × 106 cells/sample were treated with: Complete™ protease inhibitor tablets; either 1× (1 tablet for every 50 mL of media) or 2× (2 tablets for every 50 mL of media); or treated with antipain (2.0 μg/mL) plus leupeptin (2.0 μg/mL) (both from Sigma-Aldrich, St Louis, MO, USA) for cysteine protease inhibition; or aprotinin (2.0 μg/mL) plus PMSF (100 μg/mL) (both from Sigma-Aldrich) for serine protease inhibition; for to 24 hrs. ..

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  • 99
    Millipore atm kinase buffer
    Cdk5 regulates CPT-induced the re-entry of cell cycle by postmitotic neurons ( a ) Kinetics of CPT-induced activation of <t>Cdks</t> and <t>ATM.</t> CGNs were treated with 10 μM CPT for the indicated periods of time. Cdk5, ATM, Cdk2 and Cdk6 kinase activities were measured following immunoprecipitation. The lower graph is the quantification of various kinase activities. Data were shown as mean ± SD (n = 3. All Ns represent independent experiments throughout). ( b ) Phosphorylation of ATM by Cdks in vitro . Purified Cdk2, 5, and 6 were incubated with the recombinant GST-ATM4 protein in vitro in the presence of [γ- 32 P]-ATP under manufacturer’s recommended conditions (top panel) or with the standard control substrate Histone H1 (bottom panel). ( c ) The effects of dominant negative (dn) Cdks on CPT-induced γ-H2AX foci formation. CGNs were transfected with constructs for GFP and dnCdks. After 24 hours, cells were treated with 10 μM CPT for 1 hour. γ-H2AX (red) and GFP (green) were detected by immunocytochemistry, and nuclei were labeled with Hoechst (blue). The scale bar represents 5 μM. The average number of foci counted blindly are: GFP control, 0.22; CPT+GFP, 4.81; CPT+dnCdk2, 4.56; CPT+dnCdk5, 3.25, p
    Atm Kinase Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/atm kinase buffer/product/Millipore
    Average 99 stars, based on 153 article reviews
    Price from $9.99 to $1999.99
    atm kinase buffer - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Millipore m na3 vo4
    Cdk5 regulates CPT-induced the re-entry of cell cycle by postmitotic neurons ( a ) Kinetics of CPT-induced activation of <t>Cdks</t> and <t>ATM.</t> CGNs were treated with 10 μM CPT for the indicated periods of time. Cdk5, ATM, Cdk2 and Cdk6 kinase activities were measured following immunoprecipitation. The lower graph is the quantification of various kinase activities. Data were shown as mean ± SD (n = 3. All Ns represent independent experiments throughout). ( b ) Phosphorylation of ATM by Cdks in vitro . Purified Cdk2, 5, and 6 were incubated with the recombinant GST-ATM4 protein in vitro in the presence of [γ- 32 P]-ATP under manufacturer’s recommended conditions (top panel) or with the standard control substrate Histone H1 (bottom panel). ( c ) The effects of dominant negative (dn) Cdks on CPT-induced γ-H2AX foci formation. CGNs were transfected with constructs for GFP and dnCdks. After 24 hours, cells were treated with 10 μM CPT for 1 hour. γ-H2AX (red) and GFP (green) were detected by immunocytochemistry, and nuclei were labeled with Hoechst (blue). The scale bar represents 5 μM. The average number of foci counted blindly are: GFP control, 0.22; CPT+GFP, 4.81; CPT+dnCdk2, 4.56; CPT+dnCdk5, 3.25, p
    M Na3 Vo4, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m na3 vo4/product/Millipore
    Average 92 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    m na3 vo4 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

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    Cdk5 regulates CPT-induced the re-entry of cell cycle by postmitotic neurons ( a ) Kinetics of CPT-induced activation of Cdks and ATM. CGNs were treated with 10 μM CPT for the indicated periods of time. Cdk5, ATM, Cdk2 and Cdk6 kinase activities were measured following immunoprecipitation. The lower graph is the quantification of various kinase activities. Data were shown as mean ± SD (n = 3. All Ns represent independent experiments throughout). ( b ) Phosphorylation of ATM by Cdks in vitro . Purified Cdk2, 5, and 6 were incubated with the recombinant GST-ATM4 protein in vitro in the presence of [γ- 32 P]-ATP under manufacturer’s recommended conditions (top panel) or with the standard control substrate Histone H1 (bottom panel). ( c ) The effects of dominant negative (dn) Cdks on CPT-induced γ-H2AX foci formation. CGNs were transfected with constructs for GFP and dnCdks. After 24 hours, cells were treated with 10 μM CPT for 1 hour. γ-H2AX (red) and GFP (green) were detected by immunocytochemistry, and nuclei were labeled with Hoechst (blue). The scale bar represents 5 μM. The average number of foci counted blindly are: GFP control, 0.22; CPT+GFP, 4.81; CPT+dnCdk2, 4.56; CPT+dnCdk5, 3.25, p

    Journal: Nature cell biology

    Article Title: Phosphorylation of ATM by Cdk5 mediates DNA damage signaling and regulates neuronal death

    doi: 10.1038/ncb1829

    Figure Lengend Snippet: Cdk5 regulates CPT-induced the re-entry of cell cycle by postmitotic neurons ( a ) Kinetics of CPT-induced activation of Cdks and ATM. CGNs were treated with 10 μM CPT for the indicated periods of time. Cdk5, ATM, Cdk2 and Cdk6 kinase activities were measured following immunoprecipitation. The lower graph is the quantification of various kinase activities. Data were shown as mean ± SD (n = 3. All Ns represent independent experiments throughout). ( b ) Phosphorylation of ATM by Cdks in vitro . Purified Cdk2, 5, and 6 were incubated with the recombinant GST-ATM4 protein in vitro in the presence of [γ- 32 P]-ATP under manufacturer’s recommended conditions (top panel) or with the standard control substrate Histone H1 (bottom panel). ( c ) The effects of dominant negative (dn) Cdks on CPT-induced γ-H2AX foci formation. CGNs were transfected with constructs for GFP and dnCdks. After 24 hours, cells were treated with 10 μM CPT for 1 hour. γ-H2AX (red) and GFP (green) were detected by immunocytochemistry, and nuclei were labeled with Hoechst (blue). The scale bar represents 5 μM. The average number of foci counted blindly are: GFP control, 0.22; CPT+GFP, 4.81; CPT+dnCdk2, 4.56; CPT+dnCdk5, 3.25, p

    Article Snippet: To determine kinase activity of ATM or Cdks in cells, corresponding immunoprecipitates were washed twice with either ATM kinase buffer (50 mM HEPES, pH 7.5, 150 mM NaCl, 4 mM MnCl2 , 10% glycerol, 1 mM dithiothreitol, and 100 μM Na3 VO4 ) or Cdk kinase buffer, and incubated in the kinase buffer containing [γ-32 P]-ATP and 1 μg of substrate [PHAS-I for ATM kinase assay (Calbiochem) and histone H1 for Cdk kinase assay (Upstate)] for 30 min at 30°C.

    Techniques: Cycling Probe Technology, Activation Assay, Immunoprecipitation, In Vitro, Purification, Incubation, Recombinant, Dominant Negative Mutation, Transfection, Construct, Immunocytochemistry, Labeling