Compounds were tested against 20 ng of purified LdGSK-3 in the presence of 25 µM Phosphoglycogen Synthase Peptide-2 (Millipore, Ref. 12–241) and 1 µM ATP (Sigma-Aldrich, Ref. A7699) in Kinase Assay Buffer in 96-well black plates. The reaction was allowed to proceed for 30 min at 30 °C, then it was incubated at room temperature for another 10 min after addition of Kinase Glo reagents (Promega, Ref. V6712). ATP consumption by LdGSK-3 was measured indirectly through end-point luminescence measurement of each well in a Varioskan Flash microplate reader (Thermo).
Phosphoglycogen synthase peptide 2
Manufactured by Merck Group
Sourced in Germany
Phosphoglycogen Synthase Peptide-2 is a laboratory equipment product. It is a peptide that functions as a substrate for the enzyme phosphoglycogen synthase.
Automatically generated - may contain errors
Lab products found in correlation
Kinase glo reagent, by Promega (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Varioskan flash microplate reader, by Thermo Fisher Scientific (1 mentions)
Glomax96 microplate reader, by Promega (1 mentions)
Kinase glo, by Promega (1 mentions)
Adenosine triphosphate (atp), by Cell Signaling Technology (1 mentions)
2 protocols using phosphoglycogen synthase peptide 2
1
LdGSK-3 Kinase Assay Protocol
2
Kinase Activity Assay for GSK3β
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The kinase activity assay was performed as previously described [17 (link)]. Briefly, 20 ng of recombinant human GSK3β (Biomol, Hamburg, Germany) were incubated with the substrate phospho glycogen synthase peptide 2 (pGS2, 25 μM) (Millipore, Billerica, USA), ATP (1 μM) (Cell Signaling, Frankfurt am Main, Germany) and different concentrations of PDA-66 and SB-216763 for 30 min at 30°C. After addition of Kinase-Glo (Promega, Mannheim, Germany) and 10 min of incubation at room temperature the luminescence signal was measured with a Glomax 96 microplate reader (Promega).
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