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| Proteins, Expression, Isolation and Analysis | The solution pH was adjusted to pH 7.0. Then, 8?μL solubilized samples were mixed at 1:1 (v/v) ratio with 2X SDS-PAGE sample loading buffer (Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China) and were boiled for 5?min. Next, collagen samples (10?μL) were loaded onto in each well of 8% SurePAGE Bis-Tris gels (GenScript, Nanjing, China). The electrophoresis was carried out with an electrophoresis voltage of 120?V for about 50?min with a DYCZ-24KS electrophoresis cell (Beijing Liuyi Biotechnology Co., Ltd., Beijing, China) and a DYY-6D electrophoresis power supply (Beijing Liuyi Biotechnology Co., Ltd., Beijing, China). To estimate the molecular weights of proteins, broad multi-color pre-stained protein standard ranging from 5?kDa to 270?kDa (GenScript, Nanjing, China) and Spectra? multi-color high range protein ladder ranging from 40?kDa to 300?kDa (ThermoFisher Scientific, USA) were used. After electrophoresis, the gel was stained with 0.1% (w/v) Coomassie Brilliant Blue R-250 in 25% (v/v) isopropanol and 10% (v/v) acetic acid for about 3?h. Then the gel was destained using a destaining solution with 20% (v/v) ethanol and 10% (v/v) acetic acid until clear protein bands could be observed. The destained gel was put in a Gensens 1800 Gel Image Analysis System (Clinx Science Instruments Co. Ltd., China) and then the digital photo images of the destained gel were acquired by a digital camera. | Get A Quote |
Many types of collagens from different sources have been used in food, pharmaceutics, biomedicine, tissue engineering, etc. Their physicochemical properties have been widely investigated to understand their behaviors and functions. However, the polymorphism and stability of collagen?nanostructureshave not been systematically studied. In the current manuscript, polymorphism and stability of nanostructures of three types of collagens from bovine flexor tendon, rat tail, and?tilapia?skin are characterized by?sodium dodecyl sulfate-polyacrylamide gel electrophoresis?(SDS-PAGE), atomic force microscopy (AFM), and attenuated total reflectance Fourier transform infrared (ATR-FTIR)?spectrometry. SDS-PAGE results ... More
