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Method for increasing resolution power and signal strength in protein gel electrolytic zone

A technology of signal intensity and gel electrophoresis, applied in the field of protein detection, can solve the problems of weak signal, difficulty in detection and analysis, and inability to change the relative content of the target protein.

Inactive Publication Date: 2006-07-05
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, polypropylene gel electrophoresis only separates proteins in a mixture sample (such as solid organs or serum), but cannot change the relative content of a certain protein of interest
If the abundance of a certain target protein is very low, the signal of the color band of the separated protein is very weak, which will bring certain difficulties to the detection and analysis

Method used

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  • Method for increasing resolution power and signal strength in protein gel electrolytic zone

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1. Detection of Caspase-3 in mouse spleen homogenate samples.

[0022] 1. Prepare the spleen homogenate sample.

[0023] The mouse spleen was taken and homogenized in a homogenizer surrounded by ice water. Centrifuge to remove the supernatant. The supernatant sample is divided into two parts, one part is processed by the conventional method, and the other part is processed by the method of the present invention.

[0024] 2. Sample Processing

[0025] (1) Conventional treatment: the above-mentioned spleen was homogenized and the supernatant was subjected to protein concentration determination (Bradford method detection, the same below), and then a certain concentration of protein was mixed with electrophoresis loading buffer and set aside.

[0026] (2) Treatment by the method of the present invention: the supernatant of the spleen homogenate was mixed with acetone at a ratio of 1:5 (v / v), and placed at -20° C. overnight. Centrifuge (centrifugation speed is 10...

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Abstract

The invention relates to a protein detecting technical field. It is a method of strengthening the resolution ratio of the protein gel electrophoresis zone and the signal intensity. The invention includes the preparation of protein sample, the separation of the polypropylene and the colour developinent of the protein zone. It's characteristic lies in the tested protein sample is electrophoresed after treated with acetone so as to enhance the colour developing intensity to advantage the discovery of the low abundant protein so as to provide convenience for further analysis and judgment.

Description

technical field [0001] The invention relates to the technical field of protein detection, and is a method for enhancing the visibility and signal intensity of protein bands separated by gel electrophoresis in combination with antibodies against corresponding proteins and display technology. Background technique [0002] For the analysis and comparison of protein types and relative sizes of protein molecules contained in various samples such as homogenate mixture (fluid), cell lysate mixture and serum prepared from solid organs of animal or human origin, polypropylene gel electrophoresis (such as SDS -PAGE) is a commonly used technique in clinical and laboratory research. After polypropylene gel electrophoresis, the proteins in the mixture sample can be roughly separated, and then the gel is stained with Coomassie brilliant blue to roughly determine the approximate position of the protein molecule to be separated. However, polypropylene gel electrophoresis only separates pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N27/26G01N21/76G01N33/561G01N21/25
Inventor 周炳荣栾洁戚中田
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY