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PerkinElmer m egta
M Egta, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/m egta/product/PerkinElmer
Average 92 stars, based on 9 article reviews
Price from $9.99 to $1999.99
m egta - by Bioz Stars, 2020-09
92/100 stars

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In Vitro:

Article Title: Regulation of Protein Phosphatase 1I by Cdc25C-associated Kinase 1 (C-TAK1) and PFTAIRE Protein Kinase *
Article Snippet: .. For in vitro phosphorylation assays, recombinant I-2 (2 μg) or Rb (2.5 μ m ) was incubated with 50 m m imidazole (pH 7.4), 0.2 m m EGTA, 0.1% (v/v) 2-mercaptoethanol, 10 m m MgCl2 , 100 μ m ATP, and 10 μCi [γ-32 P]ATP (PerkinElmer Life Sciences) plus C-TAK1 (0.02 μg), GSK-3β (0.02 μg), C-TAK1 and GSK-3β, PFTK1 immunoprecipitate (0.2 mg of Dynabeads/reaction), CK1 (0.2 units/reaction), or CK2 (0.4 units/reaction) and incubated for 1 h at 30 °C. ..

Concentration Assay:

Article Title: Characterization of a Drosophila Ortholog of the Cdc7 Kinase
Article Snippet: .. Briefly, 25-μl reaction volumes containing a final concentration of 40 m m HEPES-NaOH, pH 7.5, 0.5 m m EDTA, 0.5 m m EGTA, 1 m m β-glycerophosphate, 1 m m NaF, 2 m m DTT, 10 m m magnesium acetate, 0.1 m m unlabeled ATP, and 2 μCi of [32 P]ATP (PerkinElmer Life Sciences, catalog no. BLU502A250UC), substrate (0.5 μg of Mcm4-GST or Mcm2-GST or 2 μg of purified HeLa core histones), and Cdc7 (6, 12, or 24 ng) or DDK (6, 12, 24 ng Cdc7 with co-purified ChiffonN) were incubated at 30 °C for 30 min (Mcm2/4 as substrate) or 60 min (core histones as substrate). .. The XL413 inhibitor was synthesized by us as described previously ( ) and solubilized in DMSO.

Purification:

Article Title: Characterization of a Drosophila Ortholog of the Cdc7 Kinase
Article Snippet: .. Briefly, 25-μl reaction volumes containing a final concentration of 40 m m HEPES-NaOH, pH 7.5, 0.5 m m EDTA, 0.5 m m EGTA, 1 m m β-glycerophosphate, 1 m m NaF, 2 m m DTT, 10 m m magnesium acetate, 0.1 m m unlabeled ATP, and 2 μCi of [32 P]ATP (PerkinElmer Life Sciences, catalog no. BLU502A250UC), substrate (0.5 μg of Mcm4-GST or Mcm2-GST or 2 μg of purified HeLa core histones), and Cdc7 (6, 12, or 24 ng) or DDK (6, 12, 24 ng Cdc7 with co-purified ChiffonN) were incubated at 30 °C for 30 min (Mcm2/4 as substrate) or 60 min (core histones as substrate). .. The XL413 inhibitor was synthesized by us as described previously ( ) and solubilized in DMSO.

Article Title: The 68-kDa Telomeric Repeat Binding Factor 1 (TRF1)-associated Protein (TAP68) Interacts with and Recruits TRF1 to the Spindle Pole during Mitosis *
Article Snippet: .. To verify whether Thr-221 is a substrate for NEK2A or Thr-457 is a substrate for PLK1, 2 μg of purified His-TAP68 fusion proteins, both wild-type and mutants, were incubated with 1 unit of active Nek2A or PLK1 (Upstate Biotechnology, Lake Placid, NY) in kinase buffer containing 20 m m HEPES, pH 7.5, 10 m m MgCl2 , 5 m m EGTA, 1 m m dithiothreitol, 10 μ m ATP, and 5 μCi of [32 P]ATP (PerkinElmer Life Sciences) in a total volume of 30 μl. .. Kinase reactions were carried out at 30 °C for 30 min and terminated by the addition of 10 μl of 4× SDS-PAGE sample buffer and separated by 6–16% gradient SDS-PAGE.

Enzyme-linked Immunosorbent Assay:

Article Title: Interferon-inducible LY6E Protein Promotes HIV-1 Infection
Article Snippet: .. Briefly, 10 μl of cell supernatants was mixed with 40 μl of reaction mixture (50 m m Tris-HCl (pH 7.9), 5 m m MgCl2 , 0.5 m m EGTA, 0.05% Triton X-100, 2% (v/v) ethylene glycol, 150 m m KCl, 5 m m DTT, 0.3 m m GSH (reduced glutathione), 0.5 units/ml poly(rA)·oligo(dT), and 0.1 μCi/μl 3 H-labeled dTTP (PerkinElmer Life Sciences)) and incubated at 37 °C for 3 h. Samples were then placed on ice for 30 min with 150 μl of prechilled 10% (mass/volume (m/v)) TCA and filtered onto an ELISA plate (Merck Millipore), and the RT activity was measured in a Microbeta counter (Beckman Coulter). .. HIV-1 infectivity was determined by infecting indicator HeLa-TZM cells.

Immunoprecipitation:

Article Title: The Phosphatidylinositol 3-Kinase/Akt Pathway Regulates Transforming Growth Factor-? Signaling by Destabilizing Ski and Inducing Smad7 *
Article Snippet: .. Immunoprecipitation with anti-HA antibody was performed as indicated above, collected on Protein G-Sepharose beads, and incubated for 30 min at 30 °C with human 100 ng of recombinant Akt (Cell Signaling Technology, Beverly, MA) in the presence of 2.5 m m β-glycerophosphate, 1 m m EGTA, 0.4 m m EDTA, 5 m m MgCl2 , 0.05 m m dithiothreitol, 25 μ m [γ-32 P]ATP (PerkinElmer Life Science) in 5 m m MOPS pH 7.2 buffer. .. Incubation was stopped by adding Laemmli buffer and boiling.

Incubation:

Article Title: Regulation of Protein Phosphatase 1I by Cdc25C-associated Kinase 1 (C-TAK1) and PFTAIRE Protein Kinase *
Article Snippet: .. For in vitro phosphorylation assays, recombinant I-2 (2 μg) or Rb (2.5 μ m ) was incubated with 50 m m imidazole (pH 7.4), 0.2 m m EGTA, 0.1% (v/v) 2-mercaptoethanol, 10 m m MgCl2 , 100 μ m ATP, and 10 μCi [γ-32 P]ATP (PerkinElmer Life Sciences) plus C-TAK1 (0.02 μg), GSK-3β (0.02 μg), C-TAK1 and GSK-3β, PFTK1 immunoprecipitate (0.2 mg of Dynabeads/reaction), CK1 (0.2 units/reaction), or CK2 (0.4 units/reaction) and incubated for 1 h at 30 °C. ..

Article Title: Characterization of a Drosophila Ortholog of the Cdc7 Kinase
Article Snippet: .. Briefly, 25-μl reaction volumes containing a final concentration of 40 m m HEPES-NaOH, pH 7.5, 0.5 m m EDTA, 0.5 m m EGTA, 1 m m β-glycerophosphate, 1 m m NaF, 2 m m DTT, 10 m m magnesium acetate, 0.1 m m unlabeled ATP, and 2 μCi of [32 P]ATP (PerkinElmer Life Sciences, catalog no. BLU502A250UC), substrate (0.5 μg of Mcm4-GST or Mcm2-GST or 2 μg of purified HeLa core histones), and Cdc7 (6, 12, or 24 ng) or DDK (6, 12, 24 ng Cdc7 with co-purified ChiffonN) were incubated at 30 °C for 30 min (Mcm2/4 as substrate) or 60 min (core histones as substrate). .. The XL413 inhibitor was synthesized by us as described previously ( ) and solubilized in DMSO.

Article Title: Interferon-inducible LY6E Protein Promotes HIV-1 Infection
Article Snippet: .. Briefly, 10 μl of cell supernatants was mixed with 40 μl of reaction mixture (50 m m Tris-HCl (pH 7.9), 5 m m MgCl2 , 0.5 m m EGTA, 0.05% Triton X-100, 2% (v/v) ethylene glycol, 150 m m KCl, 5 m m DTT, 0.3 m m GSH (reduced glutathione), 0.5 units/ml poly(rA)·oligo(dT), and 0.1 μCi/μl 3 H-labeled dTTP (PerkinElmer Life Sciences)) and incubated at 37 °C for 3 h. Samples were then placed on ice for 30 min with 150 μl of prechilled 10% (mass/volume (m/v)) TCA and filtered onto an ELISA plate (Merck Millipore), and the RT activity was measured in a Microbeta counter (Beckman Coulter). .. HIV-1 infectivity was determined by infecting indicator HeLa-TZM cells.

Article Title: The 68-kDa Telomeric Repeat Binding Factor 1 (TRF1)-associated Protein (TAP68) Interacts with and Recruits TRF1 to the Spindle Pole during Mitosis *
Article Snippet: .. To verify whether Thr-221 is a substrate for NEK2A or Thr-457 is a substrate for PLK1, 2 μg of purified His-TAP68 fusion proteins, both wild-type and mutants, were incubated with 1 unit of active Nek2A or PLK1 (Upstate Biotechnology, Lake Placid, NY) in kinase buffer containing 20 m m HEPES, pH 7.5, 10 m m MgCl2 , 5 m m EGTA, 1 m m dithiothreitol, 10 μ m ATP, and 5 μCi of [32 P]ATP (PerkinElmer Life Sciences) in a total volume of 30 μl. .. Kinase reactions were carried out at 30 °C for 30 min and terminated by the addition of 10 μl of 4× SDS-PAGE sample buffer and separated by 6–16% gradient SDS-PAGE.

Article Title: The Phosphatidylinositol 3-Kinase/Akt Pathway Regulates Transforming Growth Factor-? Signaling by Destabilizing Ski and Inducing Smad7 *
Article Snippet: .. Immunoprecipitation with anti-HA antibody was performed as indicated above, collected on Protein G-Sepharose beads, and incubated for 30 min at 30 °C with human 100 ng of recombinant Akt (Cell Signaling Technology, Beverly, MA) in the presence of 2.5 m m β-glycerophosphate, 1 m m EGTA, 0.4 m m EDTA, 5 m m MgCl2 , 0.05 m m dithiothreitol, 25 μ m [γ-32 P]ATP (PerkinElmer Life Science) in 5 m m MOPS pH 7.2 buffer. .. Incubation was stopped by adding Laemmli buffer and boiling.

Article Title: Regulation of Sarcoplasmic Reticulum Ca2+ ATPase 2 (SERCA2) Activity by Phosphodiesterase 3A (PDE3A) in Human Myocardium
Article Snippet: .. Then portions (50 μl, 10 μg of protein) were incubated at 37 °C for 1 min in 0.4 ml of Ca2+ uptake buffer consisting of 50 m m imidazole-HCl, pH 7.0, 100 m m KCl, 6 m m MgCl2 , 10 m m NaN3 (to inhibit mitochondrial Ca2+ uptake), 10 m m potassium oxalate, 20 μ m ruthenium red (to inhibit SR Ca2+ release), 0.1 m m EGTA, 45 CaCl2 (10,000 dpm/nmol) (PerkinElmer Life Sciences, catalog #NEZ013), and unlabeled CaCl2 (0.5 μ m free Ca2+ ). .. Uptake was initiated by adding 50 m m ATP (50 μl), and reactions were terminated by filtration through 0.45-μm Millipore filters.

Activity Assay:

Article Title: Interferon-inducible LY6E Protein Promotes HIV-1 Infection
Article Snippet: .. Briefly, 10 μl of cell supernatants was mixed with 40 μl of reaction mixture (50 m m Tris-HCl (pH 7.9), 5 m m MgCl2 , 0.5 m m EGTA, 0.05% Triton X-100, 2% (v/v) ethylene glycol, 150 m m KCl, 5 m m DTT, 0.3 m m GSH (reduced glutathione), 0.5 units/ml poly(rA)·oligo(dT), and 0.1 μCi/μl 3 H-labeled dTTP (PerkinElmer Life Sciences)) and incubated at 37 °C for 3 h. Samples were then placed on ice for 30 min with 150 μl of prechilled 10% (mass/volume (m/v)) TCA and filtered onto an ELISA plate (Merck Millipore), and the RT activity was measured in a Microbeta counter (Beckman Coulter). .. HIV-1 infectivity was determined by infecting indicator HeLa-TZM cells.

Kinase Assay:

Article Title: Dependence of Leucine-rich Repeat Kinase 2 (LRRK2) Kinase Activity on Dimerization *
Article Snippet: .. Recombinant WT-LRRK2 protein, truncated through the first 970 amino acids and fused with GST on the N terminus, was purchased from Invitrogen and combined into a kinase assay buffer that included PBS, pH 7.4, 5 m m EGTA, 20 m m β-glycerol phosphate, 10 μ m ATP (0.5 μCi of [γ-32 P]ATP (PerkinElmer Life Sciences)), 20 m m MgCl2 , and varying concentrations of deoxycholate (Sigma). ..

Recombinant:

Article Title: Regulation of Protein Phosphatase 1I by Cdc25C-associated Kinase 1 (C-TAK1) and PFTAIRE Protein Kinase *
Article Snippet: .. For in vitro phosphorylation assays, recombinant I-2 (2 μg) or Rb (2.5 μ m ) was incubated with 50 m m imidazole (pH 7.4), 0.2 m m EGTA, 0.1% (v/v) 2-mercaptoethanol, 10 m m MgCl2 , 100 μ m ATP, and 10 μCi [γ-32 P]ATP (PerkinElmer Life Sciences) plus C-TAK1 (0.02 μg), GSK-3β (0.02 μg), C-TAK1 and GSK-3β, PFTK1 immunoprecipitate (0.2 mg of Dynabeads/reaction), CK1 (0.2 units/reaction), or CK2 (0.4 units/reaction) and incubated for 1 h at 30 °C. ..

Article Title: The Phosphatidylinositol 3-Kinase/Akt Pathway Regulates Transforming Growth Factor-? Signaling by Destabilizing Ski and Inducing Smad7 *
Article Snippet: .. Immunoprecipitation with anti-HA antibody was performed as indicated above, collected on Protein G-Sepharose beads, and incubated for 30 min at 30 °C with human 100 ng of recombinant Akt (Cell Signaling Technology, Beverly, MA) in the presence of 2.5 m m β-glycerophosphate, 1 m m EGTA, 0.4 m m EDTA, 5 m m MgCl2 , 0.05 m m dithiothreitol, 25 μ m [γ-32 P]ATP (PerkinElmer Life Science) in 5 m m MOPS pH 7.2 buffer. .. Incubation was stopped by adding Laemmli buffer and boiling.

Article Title: Dependence of Leucine-rich Repeat Kinase 2 (LRRK2) Kinase Activity on Dimerization *
Article Snippet: .. Recombinant WT-LRRK2 protein, truncated through the first 970 amino acids and fused with GST on the N terminus, was purchased from Invitrogen and combined into a kinase assay buffer that included PBS, pH 7.4, 5 m m EGTA, 20 m m β-glycerol phosphate, 10 μ m ATP (0.5 μCi of [γ-32 P]ATP (PerkinElmer Life Sciences)), 20 m m MgCl2 , and varying concentrations of deoxycholate (Sigma). ..

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    PerkinElmer m egta
    M Egta, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m egta/product/PerkinElmer
    Average 92 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    m egta - by Bioz Stars, 2020-09
    92/100 stars
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