FHA, FHNA, and their related analogues were synthesized as described elsewhere (31 (link)). The compounds were dissolved in dimethyl sulfoxide (DMSO) to obtain 20 mM stocks and were stored at 4°C until further use. MADTP-372 was dissolved as 10 mM stock in DMSO and used as described previously (24 (link)). Mycophenolic acid (MPA), 6-azauridine (6-au), S-adenosylhomocysteine (SAH), and guanylyl-imido-diphosphate (GIDP) were purchased from Sigma. S-Adenosylmethionine (SAM) and NAD (NAD+) were obtained from New England Biolabs. S-Adenosyl-[methyl-3H]methionine, [γ-32P]ATP, and [α-32P]GTP are products of Perkin-Elmer.
S adenosyl methyl 3h methionine
Manufactured by PerkinElmer
S-Adenosyl-[methyl-3H]methionine is a radioactively labeled compound used as a substrate in various biochemical assays. It is a methyl donor that can be utilized to measure methyltransferase activity.
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Lab products found in correlation
3 protocols using s adenosyl methyl 3h methionine
1
Synthesis and Characterization of FHA, FHNA Analogues
2
Radioactive Isotope Sourcing for Experiments
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S-Adenosyl (methyl-3H) methionine (67.3 Ci/mmol) and (α-32P) GTP (3000 Ci/mmol) were sourced from PerkinElmer.
3
In vitro Methylation Assay of NSUN2
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Reaction mixtures (50 µL) containing 0.2 nM recombinant GST-tagged NSUN2, 0.01 nM in vitro transcribed fragments of mRNA, 1 µCi of S-adenosyl [methyl-3H] methionine (0.5 μCi/μl, PerkinElmer) in reaction buffer (500 mM Tris–HCl (pH 7.5), 5 mM EDTA, 10 mM dithiothreitol, 20 mM MgCl2) and 40 units of RNase inhibitor were incubated for 60 min at 37°C, as described [21 (link)]. The 3H-labeled products were isolated using DEAE-Sephadex A-50 columns and quantitated by liquid scintillation counting (PerkinElmer). Non-isotopic methylated RNA fragments were prepared using cold SAM (Biolabs, 0991410) and in vitro transcribed RNA fragments under similar conditions.
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