Tryptic digests, HPLC profile

HPLC profiles that resulted from the in gel tryptic digest of 34 pmol (2.1 pg) of a 62 kD protein are shown in Fig. 1. At this low level there are at least 5-6 peaks present in the blank chromatogram that are similar in size to peaks... 

Figure 5. Reverse phase HPLC of in gel tryptic digests of 25 pmol transferrin (A) and 10 pmol bovine serum albumin (B) and of the corresponding digests carried out on blank sections of gels (lower profiles shown in above two figures). In each instance 90% of the digest was subjected to HPLC on a 1.0 mm ID Vydac C-18 coluttm eluted at 50 pl/min. The respective full scale deflections were 18.9 mV for panel A and 4.4 mV for panel B with 0.5 volt corresponding to an absorbance of 1.0 at 210 nm.

Figure. 2. HPLC profiles of the tryptic digests of native (1). and Arg-modified derivatives PG-1 (2) and PG-2 (3) The tryptic hydrolysates were separated by HPLC on a SynChropak RP-18 column (4.6 x 25 cm) equilibrated with 0 1% TFA and eluted with a linear gradient of 0-35% acetonitrile for 105 min...

Fig. 2. Reverse phase HPLC fractionation of a tryptic digest of Ado-P..-[ H]P1 modified PRK. Sample (1.9 nmol)was loaded onto a LiChrospher RP-18 column and eluted with a 0-70% gradient of CH CN in 0.1% trifluoroacetic acid. The top panel shows 215 nm absorbance, with the inset depicting a 20-fold scale expansion to indicate (arrow) the UV peak corresponding to the radiolabeled peptide. The bottom panel shows the H profile measured using 10% of the column effluent.

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