1. Preparation of cell antigen and tissue antigen samples Second, the preparation of other samples The bacterial antigen, the secreted supernatant sample, and the extracted purified protein sample can be directly added to the electrophoresis loading buffer, and then denatured by heating in a boiling water bath for 3-4 min. Western sample preparation essentials Vendor Catalog number product name specification Catalog price Liankebio LK-WB018 PMSF (100mM) 1 g 60 Liankebio LK-WB019 RIPA lysate 100ml 150 Liankebio LK-WB004 SDS-PAGE Protein Loading Buffer (5X) 1 ml*2 30 Read the original text: http:// Freeze-dried Vegetables into powder will not affect its nutrition. Normally heat-sensitive substances are lost at high temperatures. For example: traditional high temperature drying technology. Freeze-drying technology and sun drying, drying, spray drying and so on are drying technologies, different drying methods have different effects on the quality of products. Freeze-drying method has less damage to the product. The traditional drying method is above 0℃, while the freeze-drying technology is simply understood to sublimate the moisture of fresh food under the vacuum and low temperature environment, and retain its original nutritional composition, appearance, size and other biological characteristics. Freeze-Dried Whole Vegetables,Freeze-Dried Spinach Powder,Freeze-Dried Vegetable Powder,Freeze Dried Garlic Powder Shaanxi HuiKe Botanical Development Co.,Ltd , https://www.oasis-hk.com 1. Treatment of cell antigens The RIPA lysis buffer (protease inhibitor was added before use, the final concentration was 2 ug/ml) was lysed on ice for 30-60 min, and then inserted into an ice box for sonication. Ultrasonic intensity is not suitable for foam generation. Ultrasonic 2-3s each time, repeated 3 or 4 times, centrifuged at 12000g for 3-5min, and the supernatant is taken up for use.
2. Treatment method of tissue antigen The tissue is taken out from the animal body (the sample is not suitable for repeated freezing and thawing), and a small amount (1-2g) is placed in a glass homogenizer to be ground into a homogenate, and then transferred to a microcentrifuge tube for ultrasonication. Ultrasonic intensity is based on no foam generation. Ultrasonic 5-7s each time, repeated 5 or 6 times, then 12000r/min, centrifuged for 3-5min, and the supernatant is taken up for use.
The sample treatment solution obtained by the above two methods is also added with about 1/4 volume of electrophoresis loading buffer, and then heated in a boiling water bath for 3-4 minutes to denature the protein, and then can be used as a sample for gel electrophoresis.
Introduction to Western Sample Preparation Methods