Cell lysis method for activity detection of stem cell telomere prolongation protein

Abstract

The invention discloses a cell lysis method for activity detection of stem cell telomere prolongation protein, which comprises the following steps: (1) collecting to-be-detected cells resuspended by sodium chloride injection, centrifuging to remove supernatant, and leaving cell precipitate; (2) diluting a phenylmethylsulfonyl fluoride solution, beta-mercaptoethanol and a lysis aid by using a cell lysis buffer solution, adding the three diluted solutions into the cell lysis buffer solution, and uniformly mixing in a vortex manner to obtain a cell lysis solution; (3) adding a cell lysis solution into the cell precipitate, then carefully blowing and beating the cells up and down by using a pipette until the cells are uniformly dispersed, and splitting the cells on ice; and (4) centrifuging the lysed cells at low temperature, sucking supernate, and transferring the supernate into a new centrifuge tube to obtain a cell lysis sample. The cell lysis method provided by the invention has good cell lysis effect and telomere extension protein activity detection accuracy, and effectively reduces the telomere extension protein activity detection time.

Description

technical field [0001] The invention relates to the A61 medical field in the IPC classification, in particular to a cell lysis method for detecting the activity of telomere elongation protein of stem cells. Background technique [0002] Stem cells are a class of stem cells with high self-renewal and multi-directional differentiation potential that have attracted much attention at present. In the related research of stem cells, the research on the activity of telomere elongation proteins in stem cells occupies an important research position. Because Telomerase is an enzyme responsible for telomere elongation in stem cells, it is a basic nuclear protein reverse transcriptase, which can effectively increase the number of stem cell divisions, as a hot research topic in recent years. A protein enzyme that has received more and more attention. [0003] Usually, when detecting the activity of human stem cell telomere elongation protein, stem cells need to be lysed first, but in th...

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Problems solved by technology

[0003] Generally, when detecting the activity of human stem cell telomere elongation protein, stem cells need to be lysed first, but in the prior art, for example, in the prior art (CN201911126853.9), the stem cells are lysed by simple ultrasonic shock, and the cell lysis process is for the target Ineffective simultaneous retention of nucleic acid and protein portions of telomere elongating proteins

In addition, although some commonly used phenylmethylsulfonyl fluorides have excellent protective effects on telomere extension proteins, their instability in solution also greatly affects the time period of stem cell lysis, thereby affecting telomere extension. Time and Accuracy of Protein Activity Assays

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