Reagents and equipment:
1. OptiPrepTM (600 g of iodixanol, Ï = 1.32 g/ml);
2. OptiPrepTM diluent (OD): 150mmol / L KCl, 30mmol / L MgCl2, 120mmol / L trimethylglycine - KOH, pH 7.8;
3. Homogenization medium (HM): 0.25 mol/L sucrose, 25 mmol/L KCl, 5 mmol/L MgCl2, 20 mmol/L trimethylglycine-KOH, pH 7.8; as required to OptiPrepTM diluent and homogenate medium Add protease inhibitor
4. High torque bridge motor (semiconductor switch control);
5. Potter-Elvehjem homogenizer, 20-40 ml, pores of about 0.07 mm;
6. High-speed centrifuge with a bucket rotor that can accommodate 15-50ml centrifuge tubes;
7. Syringe with metal trocar (inner diameter approx. 0.8mm);
8. Nylon filter;
experimental method:
All operations were carried out at 0-4 °C.
1. Prepare 500g/L iodixanol working solution: dip each 5-fold volume of OptiPrepTM with 1 volume of OptiPrepTM diluent;
2. Gradient solution: Diluted 500g/L iodixanol working solution with homogenizing medium (6 times volume + 4 volumes and 7 volumes + 3 volumes) to prepare two 300g / L and 350g / L An iodixanol gradient solution. Keep it on ice.
3. Remove the liver and cut into tiny pieces with scissors;
4. Transfer approximately half of the liver to a 20 ml homogenization medium of a Potter-Elvehjem homogenizer and grind it with a mortar for about 8 times and rotate at about 500 r/min to break the tissue;
5. Repeat the above for the other half of the liver chop;
6. Filter with a nylon filter;
7. Adjust the homogenate to 250 g / L iodixanol by mixing with an equal volume of 500 g / L iodixanol working solution;
8. Transfer 10-15 ml of the homogenate into a 50 ml tube, and add 10 ml each from 300 g/L and 350 g/L of iodixanol gradient solution to the homogenate;
9. Centrifuge at 100000g for 20min;
10. The nucleus is banded at the interface of 300g/L and 350g/L iodixanol, and the nuclei are aspirated by syringe and metal trocar;
11. Dilute the suspension with 2 volumes of homogenized medium and centrifuge the cells for 10 min at 2000 g to pellet the nuclei;
Ginseng has been known as the "king of herbs", "nourishing yin, nourishing health, strengthening the root" of the best products since ancient times. It contains a variety of saponins and polysaccharides. The extract can be slowly absorbed by the skin without adverse stimulation. It can expand skin capillaries, promote skin blood circulation, increase skin nutrition, regulate skin water and oil balance, prevent skin dehydration, sclerosis, wrinkling, and suppress black by ginseng active substances. It can make skin white and smooth, enhance skin elasticity and regenerate cells. It is the best product of skin care and beauty. Dip ginseng into 50% glycerin directly, rub face with glycerin 10 days later, or fry ginseng into concentrated juice, pour a little into the wash water every day, rub face with ginseng glycerin or ginseng water, can make skin quite moist.
Functions and Indications: Dabu Yuanqi, Fumai Gutuo, Bupi Yifei, Shengjin Yangxue, Anshen Yizhi. It is used for deficiency of body, cold pulse of limbs, deficiency of spleen and food, asthma and cough of lung, thirst of body and wound, internal heat to quench thirst, deficiency of Qi and blood, chronic disease, palpitation and insomnia, impotence and palace cold.
Ginseng Extract,Ginseng Tea,Ginseng Root,Panax Ginseng
Lixian Spring Pharmaceutical Co., Ltd. , https://www.lxctyy.com