0.1 μg His6-SdeA, 1 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5) and 1 mM DTT, 5 mM MgCl2, 1 mM L-glutamate. 1 mM ATP and 1 μM CaM for 2 h at 37°C. To measure the glutamylase activity of SidJ using 14C-glutamate, 2 μg His6-SdeA and 0.5 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5), 1 mM DTT, 5 mM MgCl2, 1 μCi 14C-L-glutamate (Perkin Elmer cat# NEC290E050UC), 1 mM ATP and 1 μM CaM for 2 h at 37°C. Products were resolved by SDS-PAGE gel and stained with Coomassie brilliant blue. Gels were then dried and signals were detected with x-ray films with a BioMax TranScreen LE (Kodak) for 3 d in −80°C.
Nec290e050uc
Manufactured by PerkinElmer
The NEC290E050UC is a laboratory equipment product. It is a compact and energy-efficient device designed for use in laboratory settings. The product specifications and core function are not available in this response.
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Lab products found in correlation
2 protocols using nec290e050uc
1
SidJ Glutamylase Activity Assay
2
SidJ Glutamylase Activity Assay
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0.1 μg His6-SdeA, 1 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5) and 1 mM DTT, 5 mM MgCl2, 1 mM L-glutamate. 1 mM ATP and 1 μM CaM for 2 h at 37°C. To measure the glutamylase activity of SidJ using 14C-glutamate, 2 μg His6-SdeA and 0.5 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5), 1 mM DTT, 5 mM MgCl2, 1 μCi 14C-L-glutamate (Perkin Elmer cat# NEC290E050UC), 1 mM ATP and 1 μM CaM for 2 h at 37°C. Products were resolved by SDS-PAGE gel and stained with Coomassie brilliant blue. Gels were then dried and signals were detected with x-ray films with a BioMax TranScreen LE (Kodak) for 3 d in −80°C.
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