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Broad bean leaf proteomics analysis sample preparation method

A technology for proteomics and sample preparation, which is applied in the field of sample preparation for proteomic analysis of faba bean leaves, can solve problems such as no substantial work progress, and achieve the effect of low experimental operation cost, easy operation, and simple steps

Inactive Publication Date: 2016-04-13
QINGHAI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The research on protein in broad bean started relatively late, and the main research focused on protein extraction, separation, and functional properties at the food processing level, while there was only a theoretical review of proteomics, but no substantial work progress

Method used

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  • Broad bean leaf proteomics analysis sample preparation method
  • Broad bean leaf proteomics analysis sample preparation method
  • Broad bean leaf proteomics analysis sample preparation method

Examples

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Embodiment 1

[0041] Example 1 The sample preparation method for proteomics analysis of broad bean leaves of the application

[0042] The sample preparation method for proteomic analysis of broad bean leaves disclosed in this application comprises the following steps: weighing broad bean seedling leaves→liquid nitrogen grinding→extracting buffer solution→centrifuging and discarding supernatant→acetone precipitation→vacuum drying precipitation→adding lysate→centrifugation Take the supernatant → protein quantification → aliquot freezing → sample hydration → sample preparation is complete.

[0043] (1) Accurately weigh 2.0 g of fresh samples of faba bean leaves with an electronic balance.

[0044] (2) Pre-cool the mortar by placing it in a -20°C refrigerator in advance, place the fresh broad bean leaf sample weighed in step (1) in the pre-cooled mortar, freeze it quickly with liquid nitrogen, and grind it fully to a fine powder.

[0045] (3) Quickly transfer the powder ground in step (2) to a 5...

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Abstract

The invention discloses a broad bean leaf proteomics analysis sample preparation method which comprises the following steps: taking fresh broad bean seedling leaves; cooling the fresh broad bean seedling leaves through liquid nitrogen and then grinding the cooled fresh broad bean seedling leaves into powder; adding an extracting buffer solution, after precipitation, taking supernatant, carrying out centrifugation, and removing the supernatant after centrifugation so as to obtain precipitates; adding an acetone solution, carrying out standing and centrifugation, and removing the supernatant so as to obtain precipitates; drying the precipitates to obtain powder; adding lysate into the powder; centrifuging, taking the supernatant, centrifuging again and taking the supernatant as a to-be-analyzed solution; measuring the concentration of protein in the to-be-analyzed solution; calculating the volume of the protein and sub-packaging for storage; adding a protein sample hydration solution for hydration and centrifuging so as to obtain a broad bean leaf proteomics analysis sample. With the adoption of the broad bean leaf proteomics analysis sample preparation method disclosed by the invention, the blank of a two-dimensional electrophoresis technology of broad bean leaves is filled, and the technical support is provided for deeply developing broad bean proteomics. With the adoption of the technical scheme, the steps are simple, the operation is easy, the repeatability is good, and technicians can learn and master easily. Used experiment reagents are two-dimensional electrophoresis conventional reagents, expensive and rare reagents are avoided and the experimental operating cost is low.

Description

technical field [0001] The application belongs to biochemistry, and in particular relates to a sample preparation method for broad bean leaf proteomics analysis. Background technique [0002] With the full implementation and advancement of the Human Genome Project, life science has entered the post-genome era, and its main research object has been transformed into the study of functional genomics. [0003] Proteomics takes all the proteins encoded by the genome in living cells as the research object. It is the study of the characteristics of all proteins expressed by an organism at a specific time or in a specific space, including protein expression levels, post-translational modifications, protein and protein or The interaction with other biomolecules is a new method to reveal the molecular mechanism of the interaction between plants and the environment. focus on. [0004] Two-dimensional electrophoresis (two-dimensional electrophoresis, 2-DE) can perform qualitative or q...

Claims

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Application Information

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IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 李萍张雁霞刘玉皎
Owner QINGHAI UNIVERSITY
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