. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 180 M. SAIGUSA AND H IWASAKI B —I kDa 40 23 -«- 12 kDa 100 r 80 - 40 20 h abed 100 r 80 - 60 - 40 20 h I abed Figure 3. Distribution of OHSS activity in a polyacrylamide gel divided into four equal segments after SDS-PAGE. (A) The gel. Numbers at the left of the small arrows indicate the molecular masses of the standards (see Fig. 2B). The molecular mass limits of each gel segment are indicated by the large arrows. (B) Bioassay with two or three ovigerous setal segments. Open bars indicate OHSS ac
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 180 M. SAIGUSA AND H IWASAKI B —I kDa 40 23 -«- 12 kDa 100 r 80 - 40 20 h abed 100 r 80 - 60 - 40 20 h I abed Figure 3. Distribution of OHSS activity in a polyacrylamide gel divided into four equal segments after SDS-PAGE. (A) The gel. Numbers at the left of the small arrows indicate the molecular masses of the standards (see Fig. 2B). The molecular mass limits of each gel segment are indicated by the large arrows. (B) Bioassay with two or three ovigerous setal segments. Open bars indicate OHSS activity of each gel segment. Incubation periods for the bioassay: h and h. Stippled bars: controls; the ovigerous setal segments were incubated in DW for h and h. Error bars: standard deviation. nificant difference between the two experiments. Solutions maintained for longer times (100 and 120 h) decreased in activity in both experiments. Figure 4C shows the pH dependency of OHSS activity. After an assay incubation of 1 h, the pH optimum was quite broad, about Specificity of antibodies and affinity purification An antiserum raised from molecular-sieve chromatogra- phy (fractions 6 and 7) detected only two strong protein bands on SDS-PAGE, and they appeared in fractions 6-8 (Fig. 5). These immunostained protein bands clearly corre- sponded to the peak of OHSS activity in Figure 2A. The antibodies that had stained the 32-kDa and 30-kDa protein bands were purified by immunochemical affinity to determine the specificity of binding. As shown in Figure 6A, the antibodies eluted from the 32-kDa protein band on SDS-PAGE stained the 30-kDa protein as well; and the antibodies eluted from the 30-kDa band similarly stained both the 30-kDa and 32-kDa proteins. B 100- 80- S 60^ w w O 40- 20-. 0 —i— 20 —i— 40 —i— 60 —i— 80 100 Temperature ( C) 100- 80- 60- 40 -i 20-. Please note that these images are extracted from scanned page images that may have
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
