SalB-PCP was acylated with the panthetheine-activated mechanistic probe 21 using the same methods described earlier. This holo-acyl-SalB-PCP was purified away from residual CoA proteins and small molecules using a Superdex 75 prep grade gel filtration column (16 cm x 60 cm, GE Healthcare) preequilibrated with buffer (50 mM Tris, 150 mM NaCl, 5% glycerol, pH 8.0). Purified 22 or diffusible substrates (21–25 ) were incubated with SalC or a SalC variant (20 μM) for 3 h, and then subjected to LC-HRMS analysis. All intact protein LC-HRMS was performed by the UC San Diego Molecular Mass Spectrometry Facility on an Agilent 1260 Infinity Binary LC coupled with a 6230 Accurate-Mass TOFMS.
Superdex 75 prep grade gel
Manufactured by GE Healthcare
Superdex 75 prep grade gel is a size exclusion chromatography medium designed for the separation and purification of proteins, peptides, and other biomolecules. It is composed of cross-linked agarose and dextran beads with a fractionation range of 3,000-70,000 Daltons. The gel provides high resolution separation and efficient recovery of target molecules.
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Lab products found in correlation
2 protocols using superdex 75 prep grade gel
1
Mechanistic probe acylation of SalB-PCP
2
Mechanistic probe acylation of SalB-PCP
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SalB-PCP was acylated with the panthetheine-activated mechanistic probe 21 using the same methods described earlier. This holo-acyl-SalB-PCP was purified away from residual CoA proteins and small molecules using a Superdex 75 prep grade gel filtration column (16 cm x 60 cm, GE Healthcare) preequilibrated with buffer (50 mM Tris, 150 mM NaCl, 5% glycerol, pH 8.0). Purified 22 or diffusible substrates (21–25 ) were incubated with SalC or a SalC variant (20 μM) for 3 h, and then subjected to LC-HRMS analysis. All intact protein LC-HRMS was performed by the UC San Diego Molecular Mass Spectrometry Facility on an Agilent 1260 Infinity Binary LC coupled with a 6230 Accurate-Mass TOFMS.
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