Tryptic peptide sample reconstituted with water containing 0.1% formic acid was separated by a Nano Acquity UPLC system (Waters, USA).
The mCIM substituted 400 [micro]l of water for 2 ml of
tryptic soy broth, adding just 1 [micro]l of organism instead of 10 [micro]l and extending the incubation period from 2 h to 4 h [+ or-] 15 min.
Tryptic peptide samples were reconstituted in 15 [micro]L of 3% acetonitrile with 0.1% trifluoroacetic acid.
In conclusion, we demonstrated that digested
tryptic protein samples, which contain encrypted peptides, are able to exert an antiproliferative activity on neoplastic cells.
Phosphorylation sites on the detected
tryptic peptides are shown in Figure S2.
Bacterial strains were sub-cultured in
Tryptic Soy Broth (TSB, Merck, Germany) and further incubated for 48 h at 37[degrees]C.
E.tarda can be isolated on Edwardsiella Isolation Media (EIM), Brain Heart Infusion (BHI),
Tryptic Soya Agar (TSA), Xylose Lysine Deoxycholate(XLD) and MacConkey.
Characterization of alpha-casozepine, a
tryptic peptide from bovine alpha(s1)-casein with benzodiazepine-like activity.
Following alkylation, protein solutions were subjected to
tryptic digestion, by the addition of 2.5 Og of sequencing grade porcine trypsin from Promega(R).
In previous works we demonstrated that these complexes have properties that partly overlap those displayed by native ones, being also characterized by an irreversible binding that confers on these complexes a particular resistance to
tryptic digestion [11].
Isolation and identification: Samples were inoculated in
Tryptic Soy Broth (TSB) and incubated at 37C for 24 hours.