HPLC fractionation of V8 peptides from vaccinia virus protein H5R. (a) HPLC fractionation of hydrolysate after passage through a SEP-PAK column using a gradient of 0-50% acetonitrile. The fractions encompassed by the horizontal bar (region 3) were collected for further analysis. (b) Microbore HPLC fractionation of region 3 from (a). A gradient of 15-25% acetonitrile in 0.1% trifluoroacetic acid was used and the fractions encompassed by the horizontal bars (regions I and II) collected for further analysis). (c) Microbore HPLC fractionation of region II from (b). A gradient of 0-50% acetonitrile in 20 mM NaCl (in the absence of trifluoroacetic acid - i.e. at neutral pH) was used and the fractions 28 and 29 collected for sequence analysis). The direction of increasing gradient is from left to right in all three frames; the ultraviolet absorbance trace is labelled as A280, and the radioactivity trace as 32P.
Brown et al. BMC Biochemistry 2000 1:2-2 doi:10.1186/1471-2091-1-2