基于TurboID的植物蛋白邻近标记实验方法
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邝嘉怡, 李洪清, 沈文锦, 高彩吉
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Methods for TurboID-based Proximal Labeling in Plants
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Jiayi Kuang, Hongqing Li, Wenjin Shen, Caiji Gao
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表1 实验所用相关试剂配方
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Table 1 Related reagent formulations used in experiment
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| Solution | Composition | Working concentration | | Protein Extraction Buffer | Tris-HCl (pH7.5) | 50 mmol∙L-1 | | NaCl | 150 mmol∙L-1 | | Sodium deoxycholate | 0.5% (w/v) | | SDS | 0.1% (w/v) | | EDTA | 1 mmol∙L-1 | | Triton X-100 | 1% (v/v) | | DTT | 1 mmol∙L-1 | | PMSF | 1 mmol∙L-1 | | Leupeptin | 10 μg∙mL-1 | | Protease Inhibitor Cocktail | 1× | | 4×SDS Sample Buffer | Tris-HCl (pH6.8) | 200 mmol∙L-1 | | SDS | 8% (w/v) | | Glycerol | 40% (v/v) | | β-mercaptoethanol | 20% (v/v) | | Bromophenol blue | 0.1% (w/v) | 1×PBS buffer
(pH7.4) | NaH2PO4∙2H2O | 0.263 g∙L-1 | | Na2HPO4∙12H2O | 1.856 g∙L-1 | | NaCl | 8 g∙L-1 | | KCl | 0.2 g∙L-1 | | 1×PBST | 1×PBS buffer containing 0.05% Tween-20 | Biotin stock
(stored at -20°C) | Dissolve biotin in DMSO to a final concentration of 50 mmol∙L-1 |
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