Figure 1: Determination of FSH glycoform content of pituitary hFSH21 by Western blot and dual tandem Superdex 75 gel filtration. A. Isolation of hFSH immunopurified from a 55 year old male pituitary by Superdex 75 gel filtration using a single, 10 X 300 mm column. Inset. Representative Western blot using anti-hFSHβ antibody, RFSH20, performed in triplicate. B. Representative chromatogram measuring glycoform abundance using two Superdex 75 columns combined in series and monitoring the effluent at 210 nm. C. Representative chromatogram measuring FSH glycoform abundance on the same columns, monitoring absorbance at 280 nm. The percentages in each panel indicate the mean of three determinations of the partially glycosylated hFSH21 variant relative abundance.