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. 2017 May 18;8(5):e2801. doi: 10.1038/cddis.2017.208

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Copyright © 2017 The Author(s)

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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The purification and identification of NGF from cobra venom. (a) The schemes of NGF separation from crude Chinese cobra venom. (b) Chromatogram of crude venom on a Sephadex G-75 column. Sample: 2.0 g of crude Chinese cobra venom. Column: 150 cm × 3.5 cm i.d. Elution buffer: 1% HAc. Flow-rate: 2 ml/min. The left vertical axes show: UV adsorption at 280 nm (AU). Symbols 1, 2, 3, 4 and 5 separately denote the isolated chromatographic peaks. (c) Chromatogram of the NGF fraction on a CM Sepharose CL-6B column. Sample: 30 ml of peak 5 collected from the Sephadex G-75 column. Column: 20 cm × 1.0 cm i.d. Buffer: 0.05 M NaAc-HAc (pH 5.0). Flow-rate: 1.5 ml/min. The left vertical axes show: UV adsorption at 280 nm (AU). Symbols 1 and 2 separately denote the isolated chromatographic peaks. (d) Chromatogram of the TSK-G2000-SW column. Sample: 50 μl of peak 2 collected from the CM Sepharose CL-6B column. Column: 600 mm × 7.5 mm i.d. Buffer: 0.1% TFA-0.25 M NaCl. Flow-rate: 1 ml/min. The left vertical axes show: UV adsorption at 280 nm (AU). (e) SDS-page of the fractions collected from the CM Sepharose CL-6B and TSK-G2000-SW columns. Lane 1: molecular weight marker. Lane 2: fraction containing peak 2 in c. Lane 3: fraction from the TSK-G2000-SW column. (f) Western blots was used to identify NGF and further determine the molecular weight of NGF. Lane 1: molecular weight marker. Lane 2: crude venom. Lane 3: the fractions collected from the TSK-G2000-SW column. (g and h) Effect of NGF on 8-day-old chick dorsal root ganglia. Microphotographs of the ganglia treated with the optimum concentration of NGF, 10 ng/ml (g) and 0 ng/ml NGF (h)