Amino acyl tRNA synthetases or aaRSs play a key role in assuring the precision of protein
13 translation. They are highly specific for their cognate amino acid and cognate tRNA
14 substrates during protein synthesis, utilizing ATP to ensure that proper assignments are made
15 between amino acid and anticodon. Specific aaRS for each amino acid are present in all cells.
16 We describe a new zymography technique to qualitatively visualize and semi-quantitatively
17 determine the amino acid activation capacity of each type of aaRS molecule by indirect
18 colorimetric detection of released pyrophosphates during the formation of aminoacyl-AMP.
19 Protein samples containing aaRS are subjected to Native PAGE, followed by incubation in
20 buffer containing cognate amino acid and ATP for sufficient time to generate pyrophosphates
21 (PPi) which are then converted to inorganic phosphates by pyrophosphatase treatment.