| Popis: |
A high-pressure liquid chromatographic method is described for separating the high mobility group (HMG) proteins on a 5-μm Nucleosil C18 column with the use of the ion pairing agent trifluoroacetic acid (TFA) in the mobile phase. With a multistep acetonitrile-TFA gradient, the calf thymus HMG proteins elute from this column as separate peaks in the order HMG 17, 14, 2, and 1. Protein elution is monitored by measuring the absorbance at 214 nm in a 3-mm flow cell, and the identity and purity of the peaks are confirmed by polyacrylamide gel electrophoresis and amino acid analysis. Both HMG 1 and 2 elute as multiple peaks, suggesting the presence of major variants of these two proteins. Other peaks in the chromatogram include degradation products of HMG 1 and histone H1. |
