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Pk ldh enzyme

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The PK/LDH enzyme is a lab equipment product developed by Merck Group. It is used to measure the activity of two key enzymes, pyruvate kinase (PK) and lactate dehydrogenase (LDH), which play important roles in cellular energy metabolism. The product provides a reliable and efficient tool for researchers and scientists to study these enzymes and their functions in various biological systems.

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Lab products found in correlation

Prism software, by GraphPad (2 mentions) Pep 2 phosphonooxy 2 propenoic acid, by Merck Group (2 mentions) Poly e4y, by Merck Group (2 mentions) Uv 1601 spectrophotometer, by Shimadzu (1 mentions)

Spelling variants of this product

PK/LDH (17 citations) LDH/PK enzyme mix (3 citations) LDH enzyme (2 citations)

3 protocols using pk ldh enzyme

1

ATPase Activity Assay for LE392Δuncic Mutant

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7.5 µg IOVs prepared from LE392ΔuncIC cells were incubated in 1 ml reaction containing 50 mM MOPS, pH 7.5, 1 mM dithiothreitol, 10 µL PK/LDH enzyme (Sigma), 5 mM ATP, 0.25 mM NADH, and 1.25 mM phosphor(enol)pyruvic acid at 37 °C for 10 min [16 (link)]. The reaction was started by the addition of 2.5mM MgCl2. The optical density at 340 nm was monitored for 10 min using the Shimadzu UV1601 spectrophotometer. The slope of the linear portion of each curve (between 200 and 400 s) was used to calculate ATPase activity. The relative ATPase activity was calculated as activity of mutant/WT.
Zhang H., Rahman S., Li W., Fu G, & Kaur P. (2015). Characterization of a novel domain ‘GATE’ in the ABC protein DrrA and its role in drug efflux by the DrrAB complex. Biochemical and biophysical research communications, 459(1), 148-153.
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2

Kinetic and Inhibition Assay for EGFR

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For in vitro enzyme kinetic and inhibition studies, wild type or mutant EGFR kinases (residues 696-1022 of the human EGFR) were expressed and purified as GST-fusion proteins in Sf9 insect cells essentially as described55 . Kinetic parameters were determined using the ATP/NADH coupled assay system in a 96-well plate, as described previously 55 . The reaction mixture contained 0.5 mg/mL BSA, 2 mM MnCl2, 1 mM PEP (2-(Phosphonooxy)-2-propenoic acid, Sigma-Aldrich, Cat. P-7002), 1 mM TCEP, 0.1 M HEPES (pH7.5), 2.5 mM tyrosine kinase substrate poly[E4Y] (P7244, Sigma-Aldrich), 1/50 of the final reaction mixture volume of PK/LDH enzyme (Pyruvate Kinase/Lactic Dehydrogenase enzymes from rabbit muscle, Cat. P-0294, Sigma-Aldrich), 0.5 mM NADH and 0.1~0.5 μM EGFR (0.1 μM for L858R, 0.5 μM for WT and 0.2 μM for T790M/V948R). 1 mM ATP was added last to start the reactions. Steady state initial velocity data were drawn from the slopes of the A340 curves and fit to the Michaelis-Menten equation to determine Vm and Km values. EGFR inhibition by Mig6 was measured using the same assay and Kiapp values were determined by fitting inhibition curves to the Morrison equation using Graphpad PRISM software.
Park E., Kim N., Ficarro S.B., Zhang Y., Lee B.I., Cho A., Kim K., Park A.K., Park W.Y., Murray B., Meyerson M., Beroukhim R., Marto J.A., Cho J, & Eck M.J. (2015). Structure and mechanism of activity-based inhibition of the EGF-Receptor by Mig6. Nature structural & molecular biology, 22(9), 703-711.
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3

Kinetic and Inhibition Assay for EGFR

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For in vitro enzyme kinetic and inhibition studies, wild type or mutant EGFR kinases (residues 696-1022 of the human EGFR) were expressed and purified as GST-fusion proteins in Sf9 insect cells essentially as described55 . Kinetic parameters were determined using the ATP/NADH coupled assay system in a 96-well plate, as described previously 55 . The reaction mixture contained 0.5 mg/mL BSA, 2 mM MnCl2, 1 mM PEP (2-(Phosphonooxy)-2-propenoic acid, Sigma-Aldrich, Cat. P-7002), 1 mM TCEP, 0.1 M HEPES (pH7.5), 2.5 mM tyrosine kinase substrate poly[E4Y] (P7244, Sigma-Aldrich), 1/50 of the final reaction mixture volume of PK/LDH enzyme (Pyruvate Kinase/Lactic Dehydrogenase enzymes from rabbit muscle, Cat. P-0294, Sigma-Aldrich), 0.5 mM NADH and 0.1~0.5 μM EGFR (0.1 μM for L858R, 0.5 μM for WT and 0.2 μM for T790M/V948R). 1 mM ATP was added last to start the reactions. Steady state initial velocity data were drawn from the slopes of the A340 curves and fit to the Michaelis-Menten equation to determine Vm and Km values. EGFR inhibition by Mig6 was measured using the same assay and Kiapp values were determined by fitting inhibition curves to the Morrison equation using Graphpad PRISM software.
Park E., Kim N., Ficarro S.B., Zhang Y., Lee B.I., Cho A., Kim K., Park A.K., Park W.Y., Murray B., Meyerson M., Beroukhim R., Marto J.A., Cho J, & Eck M.J. (2015). Structure and mechanism of activity-based inhibition of the EGF-Receptor by Mig6. Nature structural & molecular biology, 22(9), 703-711.
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