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Protein crystallization teaching kit and method thereof

A protein crystallization and kit technology, applied in the field of protein crystallization, can solve the problems of lack of protein crystallization experimental operation procedures, incompatibility, guidance, etc., and achieve the effects of improving experimental efficiency, reducing work intensity and saving costs.

Inactive Publication Date: 2016-07-27
HUAIHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are a variety of kits for screening protein crystallization conditions on the market, which are convenient, fast, efficient, and accurate and reliable. However, they are mainly used in scientific research and other fields, and are not suitable for basic teaching experiments.
The existing protein crystallization demonstration reagent launched by HamptonResearch mainly carries out protein crystallization experiments by purifying lysozyme in egg white. This demonstration reagent provides convenience for scientific researchers to carry out protein crystallization technology training, but there are still the following disadvantages: It is not suitable for basic biological science teaching experiments: (1) The demonstration reagent is not a whole kit, and the reagents and materials needed for the experiment need to be purchased separately; (2) The kit lacks specific operations for protein crystallization experiments The guidance of the process is not easy for students to understand; (3) It is impossible to distinguish between salt crystals and protein crystals

Method used

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  • Protein crystallization teaching kit and method thereof
  • Protein crystallization teaching kit and method thereof
  • Protein crystallization teaching kit and method thereof

Examples

Experimental program
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Embodiment 1

[0036] Embodiment 1, a teaching kit for protein crystallization, which consists of the following items:

[0037] (1) Protein precipitant A, 1 bottle, its composition is: PEG4000 with a volume fraction of 30%, sodium acetate buffer solution with a concentration of 0.05M and pH4.5, and sodium chloride with a concentration of 1M;

[0038] (2) Protein precipitant B, 1 bottle, which is composed of: PEG4000 with a volume fraction of 4%, sodium acetate buffer solution with a concentration of 676.7mM and pH3.0, and sodium chloride with a mass concentration of 13.3%;

[0039] (3) Protein precipitant C, 1 bottle, its composition is: sodium acetate buffer solution with a concentration of 0.1M and pH 7.4, and sodium chloride with a mass concentration of 6.5%;

[0040] (4) Lysozyme powder, 20mg, 1 bag;

[0041] (5) Dilution buffer: sodium acetate buffer solution with a concentration of 0.02M and pH 4.6, 1 bottle;

[0042] (6) Dyeing agent: methylene blue with a mass concentration of 0.3%...

Embodiment 2

[0046] Embodiment 2, the teaching kit of protein crystallization described in embodiment 1, described lysozyme powder is egg white lysozyme powder.

Embodiment 3

[0047] Embodiment 3, using the teaching kit described in embodiment 1 or 2 to carry out the method for protein crystallization, its steps are as follows:

[0048] (1) Prepare lysozyme powder with dilution buffer to make a lysozyme solution with a final concentration of 10 mg / mL, filter through a filter to remove impurities, and set aside;

[0049](2) Place the lid of the crystallization plate upside down on the table, take 5 μL of the lysozyme solution and protein precipitation agent A or B obtained in step (1) according to the volume ratio of 1:1 and drop them on the center of the crystallization plate cover, and then add 1 μL dropwise for staining Add 2mL of the above-mentioned protein precipitation agent to the crystallization plate, and carefully cover the crystallization plate cover on the crystallization plate to prevent the droplets from dripping;

[0050] (3) Seal the crystallization plate with parafilm, let it stand at room temperature for 20-30min, and then observe t...

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Abstract

The invention discloses a protein crystallization teaching kit. Articles in the kit include the following: three different protein precipitants, lysozyme powder, a dilution buffer, a dye, a filter, a crystallization plate and a sealing film. The invention also relates to a protein crystallization method which makes use of the teaching kit. The teaching kit disclosed by the invention, which contains reagents and materials required by a lysozyme crystallization experiment in biological science teaching therein, can reduce the working intensity of experiment preparation, and the teaching kit is simple and convenient to operate, conducive to experiment personnel to master and complete a protein crystallization experiment within a relatively short time, and capable of improving an experimental efficiency and saving cost; therefore, the teaching kit can be widely popularized in biological basic teaching experiments.

Description

technical field [0001] The invention relates to a teaching kit for protein crystallization, which is used for protein crystallization experiments in biological science teaching. The present invention also relates to a method for protein crystallization using the aforementioned kit. Background technique [0002] The three-dimensional structure of proteins is extremely important information in life science research. The most commonly used method for analyzing the three-dimensional structure of proteins is X-ray diffraction, and obtaining high-quality protein crystals is an important prerequisite for protein structure analysis using this technology. Egg white lysozyme is a protein that is easier to grow into an ideal single crystal. It has the characteristics of simple crystallization reagents, short crystallization time, and easy growth of protein crystals. Therefore, it is used as an important model protein and is widely used in protein crystallization mechanisms and Methods...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G09B23/26C07K1/30
CPCG09B23/26C07K1/306
Inventor 许恒皓张坤晓李婷婷韩挺翰程艺陈凯
Owner HUAIHAI INST OF TECH
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