生化实验步骤.docVIP

一、Purpose 1.In order to understand the function of differential expression proteins in leaves of seedlings, the proteins Of leaves at the seedling stage of hybrid Shanyou63 were studied by using approach of plant proteomics. 2.In order to learning the methods of Two-Dimensional Electrophoresis, TCA/acetone method to extract proteins, The methods of SDS. 3. Familiar with the operation and equipment in the laboratory. 4. Familiar with high performance liquid instruments 5 Familiar with the flow and operation method of HPLC 二、Principles 1. TCA method TCA method is also called precipitation method refers to the cohesion protein molecules from the solution .This phenomenon is known as protein precipitation 2. 2-DE A direction to SDS-polyacrylamide gel to separate proteins in accordance with the molecular weight ,A direction to IEFF isoelecctric focusing elecctrophoresis to separate the different proteins in accordance with the isoelectric point . So that can separate the different proteins 3 .HPLC High-performance liquid chromatography (sometimes referred to as high-pressure liquid chromatography), HPLC, is a chromatographic technique used to separate a mixture of compounds in analytical chemistry and biochemistry with the purpose of identifying, quantifying and purifying the individual components of the mixture. Some common examples are the separation and quantitation of performance enhancement drugs (e.g. steroids) in urine samples, or of vitamin D levels in serum. HPLC typically utilizes different types of stationary phases (i.e. sorbents) contained in columns, a pump that moves the mobile phase and sample components through the column, and a detector capable of providing characteristic retention times for the sample components and area counts reflecting the amount of each analyte passing through the detector. The detector may also provide additional information related to the analyte, (i.e. UV/Vis spectroscopic data, if so equipped). Analyte retention time varies d