Easytag l 35s met

The RRL supplemented with [³⁵S]-Met (EasyTag™ L-[³⁵S]-Met; 10 mCi/mL; PerkinElmer, Waltham, United States) was used for in vitro translations (Pelham and Jackson, 1976 (link)) of in vitro transcribed anxA2 mRNA. AnxA2 was added in the same buffer as described above and the RRL constituted 70% of the assays. Incorporation of [³⁵S]-Met into AnxA2 protein was measured as described above.

The TNT^® T7 Quick for PCR DNA (Promega, Madison, United States) is an in vitro coupled transcription/translation system based on rabbit reticulocyte lysate (RRL) and was supplemented with the cDNA coding for full-length rat anxA2 mRNA (including the UTRs and containing a T7 promoter site; described above) and [³⁵S]-Met (EasyTag™ L-[³⁵S]-Met; 10 mCi/mL; PerkinElmer, Waltham, United States). AnxA2 in 20 mM Tris (pH 8) was added to the RRL before the addition of the cDNA and the RRL constituted 63% of the assays. The reaction was performed as previously described (Strand et al., 2021 (link)). Incorporated [³⁵S]-Met into the AnxA2 protein and total radioactivity in the reaction mixture were measured in a Packard liquid scintillation counter.