. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 176 D. E. MORSE ET AL hydrophobic interaction chromatographic resin. The morphogenic inducer retains its activity after adsorption to these hydrophobic supports and is not eluted by sea- water (although penetrance into the pores of the nitro- cellulose niters apparently reduces detectability by the larvae, requiring that the niters be minced before assay). The adsorbed inducer can be subsequently released from the HIC support by elution with low ionic strength buffer (Fig. 1). This chromatographic adsorption and subsequent e
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 176 D. E. MORSE ET AL hydrophobic interaction chromatographic resin. The morphogenic inducer retains its activity after adsorption to these hydrophobic supports and is not eluted by sea- water (although penetrance into the pores of the nitro- cellulose niters apparently reduces detectability by the larvae, requiring that the niters be minced before assay). The adsorbed inducer can be subsequently released from the HIC support by elution with low ionic strength buffer (Fig. 1). This chromatographic adsorption and subsequent elution concentrated the active morphogen by about 100- fold from the dilute demineralized crude extract (per- mitting direct assays of the morphogen in solution), re- sulting in significant purification. As seen in Figure 1, the peak of morphogenic activity is eluted in fractions 2-5, whereas the majority of the light-absorbing contaminants are eluted in the first fraction. At the lower concentrations assayed, activity of the purified inducer is approximately proportional to the amount added (Fig. 2). Induction of metamorphosis is both time- and dose-dependent, with metamorphosis observed as quickly as 2-3 h following addition of high concentrations of the HIC-purified in- ducer. 100 -. 1 2 3 4567 Fraction 8 9 Figure 1. Hydrophobic-interaction chromatography. The clarified demineralized nitrate (4 1) was adjusted to 2 M NaCl, adsorbed to 8 g of the t-butyl HIC resin, poured into a column, and eluted with Tris Cl buffer as described in Materials and Methods. Fractions of 12 ml were collected; ml of each fraction was assayed for morphogenic activity (solid line). Absorbance at 212 nm is shown (dashed line).. Please note that these images are extracted from scanned page images that may have been digitally enhanced for readability - coloration and appearance of these illustrations may not perfectly resemble the original Marine Biological Laboratory (Woods Hole, Mass. );
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
