Method for storing protein sample for proteomics at normal temperature
A technology for proteomics and normal temperature storage, applied in the preparation of test samples, etc., can solve the problems of difficult protein storage and transportation, inconvenient transportation, etc., and achieve the effects of easy transportation, good mass spectrometry compatibility, and good quality
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Embodiment 1
[0025] Bovine serum albumin (BSA) was purchased from BIOSHARP Company, and the powder of bovine serum albumin (BSA) was dissolved in Tris-saturated phenol (pH 8.0), and Tris-saturated phenol (pH 8.0) was purchased from Beijing Suolaibao Technology Co., Ltd. company, and then add the phenol phase to 5 times the volume of supersaturated ammonium sulfate methanol solution (ammonium sulfate is added to the methanol solution to make the solution supersaturated, and put it at 4°C for overnight use) to precipitate the protein, and the precipitated protein sample They were placed in a supersaturated ammonium sulfate methanol solution in a 30°C incubator for 0 days, 1 day, 3 days, 5 days, 10 days and 15 days, and the final samples were placed at -20°C for use. It should be noted that the supersaturated ammonium sulfate methanol solution is supersaturated at any temperature: after being prepared at room temperature, store it in a refrigerator at around 4°C, and take it out before use.
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Embodiment 2
[0031] The rubber tree latex C-whey protein crude extract dissolved in Tris-saturated phenol with a pH value of 8.0 extracted according to the method in Example 1, after precipitation with 6 times the volume of supersaturated ammonium sulfate methanol solution, the precipitate The resulting protein was placed in a supersaturated ammonium sulfate methanol solution in a 30°C incubator for 0 days, 1 day, 3 days, 5 days, 10 days and 15 days, and the lysed protein was washed according to the method in Example 1 to obtain 800 micrograms C-The protein in whey was hydrated in a hydration dish for 18 hours with a 24 cm long linear IPG gel strip (GE Healthcare) with a pH value of 4-7, and then the isoelectric focusing process was completed using a focuser (Hoefer IEF100), The strip after completion was firstly placed in equilibrium solution (50mM Tris-HCl, pH8.8, 6M urea, 30% glycerol by volume, 2% sodium dodecylsulfonate (SDS), mass percent Percentage content is 0.02% bromophenol blue)...
Embodiment 3
[0044] According to the BPP method (Wang et al.2007) in Example 1, extract the protein in sugarcane leaves and rubber tree bark, after precipitating proteins with 4 times the volume of supersaturated ammonium sulfate methanol solution, a part of the samples of sugarcane leaves are placed in -20 Preserve in supersaturated ammonium sulfate methanol solution for two months, and another part of sugarcane leaf sample carries out protein washing and cracking according to the method in Example 1 (not preserved in supersaturated ammonium sulfate methanol solution), in 4 ℃ refrigerator stored for two months. The samples stored in supersaturated ammonium sulfate methanol solution at -20°C were also washed and lysed according to the method in Example 1. Two-dimensional electrophoresis experiments were carried out on samples with two different storage methods, and the results are shown in image 3 . It can be seen from the two-dimensional electrophoresis pattern that after the sample is...
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