A new method for the determination of intracellular protein content normalization

A technology of protein content and new method, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of easy external influence and complicated operation.

Active Publication Date: 2017-10-24
河北北科生物科技有限公司
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Problems solved by technology

Standardization with total protein has disadvantages such as the need to lyse cells, complicated operations, and the results are easily affected by the outside world.

Method used

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  • A new method for the determination of intracellular protein content normalization
  • A new method for the determination of intracellular protein content normalization

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Embodiment Construction

[0014] The present invention will be further described below:

[0015] The present invention comprises the following steps:

[0016] (1) Preparation of the standard curve: Take the cell solution of known concentration, dilute it by 2 times, 4 times, 6 times, 8 times, 10 times, etc., and then take the same amount of cell suspension for DNA fluorescent staining. Use a fluorescence spectrophotometer to measure the relative content of fluorescence, take the cell concentration as the abscissa, and the measured fluorescence value as the ordinate, and draw the standard curve;

[0017] (2) Determination of the sample to be tested: Take an equal amount of the sample to be tested and carry out DNA fluorescent staining. The operation method is the same as the preparation of the standard curve. Determination, the measured fluorescence value can find out the concentration of cells from the standard curve;

[0018] (3) The standard protein content of the sample is calculated according to ...

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Abstract

The invention discloses a new standard method for measuring intracellular protein content, which is mainly achieved by preparing a standard curve and measuring samples to be tested, and adopts DNA fluorescent staining. DNA is the genetic material in cells, and different For cells, the DNA content is relatively stable, and using DNA to standardize the protein to be tested can overcome many shortcomings of protein standardization.

Description

technical field [0001] The invention relates to a method for measuring the quantity of biological cells, in particular to a new method for measuring the standardized content of intracellular protein. Background technique [0002] When measuring the content of a certain protein (or enzyme activity) in cells, in order to compare different samples, standardization processing is required to compare different samples. For example, the activity of intracellular succinate dehydrogenase in two samples A and B is determined. When we measure the activity of intracellular succinate dehydrogenase in samples A and B by a certain method, we cannot directly conclude which sample has the higher activity of the enzyme, because the number of cells in the two samples may be different. Differential, therefore, requires normalization of enzyme activity across samples. [0003] At present, to standardize the activity of enzymes in different samples, the commonly used method is to measure the to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 井长勤于毅任太芳张业宏解丽芹包小丹
Owner 河北北科生物科技有限公司
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