Safety is the premise of experimentation; standardized procedures are the foundation of experimentation.

Release time:

2021-03-12

Overview:

An Explanation of the Application of Centrifuges in Protein Extraction from Plant Tissues. The extraction method described in this article employs the trichloroacetic acid-acetone precipitation method. The detailed steps are as follows:

　　An explanation of the application of centrifuges in protein extraction from plant tissues.

　　The extraction method described in this article employs the trichloroacetic acid-acetone precipitation method. The detailed steps are as follows:

　　1. Grind the leaves in liquid nitrogen.

　　2. Add an extraction solvent equal to three times the volume of the sample, and incubate overnight at -20℃. Then, centrifuge the mixture (at 4℃ at 8,000 rpm or higher for 1 hour), and discard the supernatant.

　　3. Add an equal volume of acetone chilled in an ice bath (containing 0.07% β-mercaptoethanol), mix thoroughly, and then centrifuge at 4°C at over 8,000 rpm for 1 hour. Subsequently, vacuum-dry the precipitate and set it aside for future use.

　　4. Add lysis buffer before sample loading, and let it stand at room temperature for 30 minutes to ensure that the protein is fully dissolved in the lysis buffer. Then, centrifuge using a centrifuge (at 15°C at 8,000 rpm or higher for 1 hour or longer, with no visible precipitate as the criterion). The sample can be temporarily stored at 4°C for later use.

　　5. Quantify the protein using the Bradford assay, then aliquot and store at -80℃ for future use.

　　Medicine:

　　Extract solution: Acetone containing 10% TCA and 0.07% β-mercaptoethanol

　　Lysis buffer: Dissolve 2.7 g of urea and 0.2 g of CHAPS in 3 ml of sterile, deionized water (final volume: 5 ml). Before use, add 1 M DTT at a concentration of 65 μl/ml.

　　This method is particularly suited to two-dimensional electrophoresis, given its characteristics of few impurities and low ion concentration! Of course, it’s also applicable to one-dimensional electrophoresis—though in that case, the number of bands will be reduced!

　　Please note that during the use of the centrifuge, certain safety requirements must be strictly followed:

　　The strength calculation of the rotating drum shall comply with the provisions specified in the scope of application of JB/T 8105. Its structure and all its components and parts must possess sufficient strength, rigidity, and stability.

　　Under the specified operating conditions, the equipment shall ensure safe and reliable operation and be easy to operate and maintain. Provided that operators comply with the operating procedures, the safety protection devices shall effectively ensure personal safety.

　　When manufactured, installed, stored, transported, and used in accordance with the specified conditions, it shall not pose a threat to or cause harm to personal safety.

　　Centrifuge Sealed and Explosion-Proof Requirements

　　1) Centrifuges operating in enclosed, flammable, explosive, or humid environments shall comply with relevant standards or regulations concerning enclosure, flame retardancy, explosion-proof measures, and moisture resistance.

　　2) Centrifuges operating in enclosed, flammable, explosive, or humid environments shall be equipped with reliable safety protection measures.

　　Shanghai Luxiangyi Centrifuge Instrument Co., Ltd., as an experienced centrifuge manufacturer, strictly adheres to the above requirements when designing and manufacturing centrifuges as well as providing training on their use!

Analysis of the Reasons for the Diversified Prices of Centrifuges

How to Meet the Special Centrifugation Requirements for Blood Banks