Elution device for protein separation and columnar gel electrophoresis device using same

An eluting device and gel electrophoresis technology, applied in the field of biochemical detection and analysis, to achieve the effect of light weight, short manufacturing process, fast and accurate on-line separation and detection

Active Publication Date: 2019-03-01
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no precedent for applying 3D printing technology to columnar gel electrophoresis devices.

Method used

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  • Elution device for protein separation and columnar gel electrophoresis device using same
  • Elution device for protein separation and columnar gel electrophoresis device using same
  • Elution device for protein separation and columnar gel electrophoresis device using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The horizontal columnar gel electrophoresis device of the present invention is used to separate the protein standard off-line, and compare with the commercialized reference results.

[0058] The filler in the columnar gel tube is 50 μl of 4% stacking gel and 400 μl of 10% separating gel, and the added protein standard is the Precision Plus Protein Dual Color protein standard produced by Bio-Rad. The buffer solution used in the electrophoresis tank is Tris-glycine-SDS buffer, and the elution solution is 50mM NH 4 NO 3solution, and separated under segmental voltage. The first section of the voltage used is 60V for 30 minutes and the second section is 200V for 5 hours. The separation results are as follows Figure 4A and Figure 4B as shown, Figure 4A It is the result of separating the Precision Plus Protein Dual Color protein standard produced by Bio-Rad Company by the columnar gel electrophoresis device of the present invention; Figure 4B SDS-PAGE separation effec...

Embodiment 2

[0060] Use horizontal columnar gel electrophoresis device of the present invention to two kinds with I 127 Labeled standard proteins were separated online and detected by ICP-MS.

[0061] The polyacrylamide gel filled in the gel tube used is 40 μl concentration 4% stacking gel and 180 μl concentration 10% separation gel. 127 The two labeled standard proteins were purchased from Sigma, USA. The sample loading volume was 20 μL, and the protein concentration was 0.01 μg / μL. The electrophoresis voltage program was 100V for 10 minutes, 200V for 20 minutes, and 600V for 2 hours. The electrophoresis buffer salt solution used is: Tris-glycine-SDS solution. Protein eluent was 50mM NH 4 NO 3 solution.

[0062] The rotational speed of the external peristaltic pump was 140 μL / min, and the ICP-MS used was 8800 ICP-MS produced by Agilent. The incident power is 1500W, the collision gas flow rate is 15.0L / min, the carrier gas flow rate is 0.80L / min, the auxiliary gas flow rate is 0.80L / m...

Embodiment 3

[0066] Such as Figure 6A , 6B Shown, use horizontal columnar gel electrophoresis device of the present invention to three kinds with I 127 The labeled standard proteins were separated online and detected in conjunction with ICP-MS, and the same standard proteins were detected using an optimized commercial column gel electrophoresis device under the same conditions, because the proteins may contain metal elements , so the I 127 and Zn 65 signal and compared the results.

[0067] The polyacrylamide gels filled in the gel tubes used are 50 μl concentration 4% stacking gel and 200 μl concentration 10% separation gel. 127 The three labeled standard proteins were purchased from Sigma, USA. The sample loading volume was 20 μL, and the protein concentration was 0.01 μg / μL. The electrophoresis voltage program was 100V for 10 minutes, 200V for 20 minutes, and 600V for 2 hours. The electrophoresis buffer salt solution used is: Tris-glycine-SDS solution. Protein eluent was 50mM NH ...

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Abstract

The invention provides an elution device for protein separation and a columnar gel electrophoresis device adopting the elution device. The elution device comprises a front part and a rear part which can be assembled; a main body part of the elution device is manufactured with a 3D printing technology. The elution device can be used for on-line separation and detection of protein with different molecular weights in combination with ICP-MS (inductively coupled plasma mass spectrometry) and other instruments, has the characteristics of short analysis time, low cost, simple manufacturing process, small dead size, wide selection range of separation conditions, high separation degree and the like, can be modified according to actual needs, and has the possibility of constant optimization.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection and analysis, and more specifically relates to an elution device for protein separation and a columnar gel electrophoresis device using the same. Background technique [0002] As the basic composition of organisms, protein is the executor of various important functions of organisms. The study of protein composition, structure and function has important research significance and practical value. Among them, electrophoresis is the most widely used and effective method in protein separation and determination. [0003] Electrophoresis techniques mainly include isoelectric focusing electrophoresis (IEF) and polyacrylamide gel electrophoresis (PAGE). PAGE uses polyacrylamide gel with a network three-dimensional structure as a supporting medium. According to the molecular charge, size and shape, it is separated into many distinct zones. PAGE can be divided into denaturing (SDS-PAGE) and n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N27/62C07K1/26
CPCC07K1/26G01N27/447G01N27/62
Inventor 何滨王丁一严雪婷胡立刚江桂斌
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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