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Shaking creates bubbles and mechanical stress that may degrade the peptide
Although L-tyrosine is a non-essential amino acid that can be synthesized endogenously from phenylalanine via the enzyme phenylalanine hydroxylase, its synthesis may become insufficient under conditions of physiological stress, rapid growth, or limited phenylalanine availability [71]
Subsequently, the blots proceeded through a second round of immunodetection by agitating for 45 min at 50C in stripping buffer (2% SDS, 0.8% -mercaptoethanol (ME), and 62.5 mM Tris-HCl, pH 6.8) 32,56,62 followed by re-probing with HRP-labeled anti-HGF N -terminal domain mAb (1:2500 dilution in CanGetSignal solution 1) to monitor HGF -chain amount on the blots as loading controls
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PMCID: PMC7432488