Cell and tissue whole protein extracting method

Abstract

The invention relates to a cell and tissue whole protein extracting method. Denaturing lysis buffer and nondenaturing lysis buffer are used as reagents respectively, and a centrifugal machine, a 1.5 ml centrifugal test tube and a 50 ml columnar polyethylene centrifugal test tube with a diaphragm in the middle act on an extracting tool. The cell and tissue whole protein extracting method comprises the following steps: step one, placing cells in the 1.5 ml centrifugal test tube, adding 1 ml of a PBS cell washing solution, then centrifuging for 3 minutes by using the centrifugal machine, removing supernate, and repeating a process of centrifuging and removing the supernate for three times; second step, adding the pre-cooled PBS cell washing solution according to the sizes of cell pellets, and carrying out cell resuspension; third step, adding the denaturing lysis buffer according to the amount of the cells in the centrifugal test tube, and oscillating for 15 seconds; fourth step, transferring the oscillated denaturing lysis buffer into the 50 ml columnar polyethylene centrifugal test tube, centrifuging for 30 seconds by using the centrifugal machine, and then collecting the lysis buffer at the bottom of the tube to obtain a whole cell protein extracting liquid; and fifth step, adding a protease inhibitor. The speed can be increased, and protein loss is reduced.

Description

technical field [0001] The invention relates to a whole protein extraction process, in particular to a cell and tissue whole protein extraction method that can rapidly and efficiently extract whole protein from cell tissues and can prevent partial protein loss caused by DNA precipitation during cell lysis. Background technique [0002] At present, the known method for extracting whole protein from cell tissue mainly adopts RIPA lysate as extraction reagent, and adopts RIPA lysate as extraction reagent to extract whole protein and has the shortcoming of slow speed, after cells are washed with PBS cell washing solution, generally It takes 20 to 30 minutes to complete all subsequent whole protein extraction processes. Due to the long extraction time, some protein will be lost due to DNA precipitation during cell lysis. Contents of the invention [0003] In order to overcome the current disadvantages of using RIPA lysate as the reagent for extracting whole protein from cell ti...

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Problems solved by technology

[0002] At present, the known method for extracting whole protein from cell tissue mainly adopts RIPA lysate as extraction reagent, and adopts RIPA lysate as extraction reagent to extract whole protein and has the shortcoming of slow speed, after cells are washed with PBS cell washing solution, generally It takes 20 to 30 minutes to complete all subsequent whole protein extraction processes. Due to the long extraction time, some protein will be lost due to DNA precipitation during cell lysis.

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