. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. FIG. 1. The Acidic PAGE of the different fractions after separation with a Sephadex G-50 column ( x 200 cm). 1: Crude venom; 2-11: Different fraction after separation with a Sephadex G- 50 column. Fractions 7-11 were collected together for the further isolation. containing NaCl gradient () and finally was eluted by the buffer containing NaCI gradient (). The two neurotoxins were eluted during the first gradient process. The neurotoxins from CM- Sephadex C- 25 still showed two other mi
. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. FIG. 1. The Acidic PAGE of the different fractions after separation with a Sephadex G-50 column ( x 200 cm). 1: Crude venom; 2-11: Different fraction after separation with a Sephadex G- 50 column. Fractions 7-11 were collected together for the further isolation. containing NaCl gradient () and finally was eluted by the buffer containing NaCI gradient (). The two neurotoxins were eluted during the first gradient process. The neurotoxins from CM- Sephadex C- 25 still showed two other minor bands on acidic PAGE. So each of them was further purified on the CM- Cellulose 52 Column. The equilibrium buffer is , pH HAc-NaAc buffer. The NaCI gradient is in the same buffer. 2. Determination of purities: Acidic PAGE was used. The separating gel was 15 %, and the spacal gel was %. The gel was stained by R- 250 dissolved in 375 ml ethanol, 125 ml water and 5 ml methanol. 3. Determination of amino acid composition: The samples were analyzed with a Hitachi Model 835-50 High Speed Amino Acid Analyzer. 4. Determination of amino acid sequence of the neurotoxin: The method was in accordance with the method of Lin and Wang (1984).. FIG. 2. The acidic PAGE of the neurotoxins from CM-Sephadex C-25 CM-Cellulose columns. 1. Crude venom; 2. The fraction containing neurotoxins from Sephadex G-50; 3. Neurotoxin 1; 4. Neurotoxin 2 from CM Sephadex C-25; 5. Neurotoxin 2 further purified on CM cellulose 52. 5. Determination of LD 50: Mice were used as the material. The method was according to Gu (1965). 6. Morphine withdrawal jumping model: Mice were divided into random groups, with each group consisting of 10 mice, half male and half female, with body weights of 20±2g. The groups were treated by injecting S. V. with morphine, for four days, three times at a dose of 10 mg/kg, three times at a dose of 20 mg/kg and 12 times at a dose of 30 mg/kg. After morphine was gi
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