Calf chondroprotein, preparation and use thereof
A cartilage protein, calf technology, applied in the preparation method, application, food preparation and other directions of peptides, can solve problems such as restricting development
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Embodiment 1
[0027] Embodiment 1: the preparation of calf cartilage protein
[0028] 10kg of newborn calf ears are peeled, washed, mashed and soaked in 1mol / L 10L acetate buffer overnight, usually for 12-16 hours. Centrifuge to remove the precipitate, take the supernatant and add 2 times the volume of saturated ammonium sulfate for fractional precipitation (the final saturation of ammonium sulfate is 50%). At 4°C for more than 3 hours, make it fully precipitate and centrifuge (3000rpm) for 20 minutes, discard the supernatant, dissolve the precipitate with 1L of normal saline, and then add 500ml of saturated ammonium sulfate dropwise. Put it at 4°C for more than 3h, [at this time, (NH4) 2 The saturation of SO4 is 33%] repeat above-mentioned step process 2 times. The precipitate obtained after the final centrifugation is protein, which is dissolved in 0.02% mol / L pH7.4 PBS to 500 ml and loaded into a dialysis bag. Fully dialyze the PBS and change the solution 3 times until the dialyzed fl...
Embodiment 2
[0029] Embodiment 2: the separation of calf cartilage protein
Embodiment 3
[0031] Embodiment 3: Calf cartilage protein electrophoresis qualitative analysis
[0032]Method reference: Sambrook et al., Molecular Cloning Test Guide, Second Edition, 1995, Science Press, China, Beijing. Weigh 10ml each of calf cartilage protein A, calf cartilage protein B, calf cartilage protein C, calf cartilage protein D, and calf cartilage protein E, and fully dissolve them in a test tube containing 1.0 ml of double distilled water. Prepare 0.02ml of sample solution. The sample and the standard protein were subjected to 12% SDS-polyacrylamide gel electrophoresis, and the electrophoresis condition was 100v for about 1 hour. After electrophoresis, the polyacrylamide gel was stained and decolorized with Coomassie Brilliant Blue R-250 dye. Comparing the obtained results with the standard proteins, the molecular weights of calf cartilage protein A, calf cartilage protein B, calf cartilage protein C, calf cartilage protein D, and calf cartilage protein E are 66Kd, 32Kd, 32K...
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