CN201068436Y

CN201068436Y - Protein condensation/purification device

Info

Publication number: CN201068436Y
Application number: CNU2007200799170U
Authority: CN
Inventors: 李胜联; 陆明深; 施文祥; 陈森洲; 仇小强
Original assignee: Guilin Medical University
Current assignee: Guilin Medical University
Priority date: 2007-06-08
Filing date: 2007-06-08
Publication date: 2008-06-04
Anticipated expiration: 2017-06-08

Abstract

The utility model provides a protein concentration/purification device, which is mainly composed of a direct-current power supply, an original pool of protein to be separated and purified, an anode pool, a cathode pool, and an isoelectric gel bridge pipe. The anode and the cathode of the direct-current power supply are respectively connected with the anode pool and the cathode pool, and the gel bridge pipe is respectively communicated with the original pool and the anode pool, and the original pool and the cathode pool. The device utilizes amphoteric ion, such as amino acid and protein in different pH solutions, the electrical charge properties of which are different, under the action of direct current field, as long as the isoelectric points of the amino acid and the protein are not equal to the pH values of the original pool solutions, the amino acid and the protein are respectively moved to the anode pool or the cathode pool by the original pool through the gel of a gel pipe, when the isoelectric points the amino acid and the protein are equal to the pH value of the original pool solution, and the protein and the amino acid which are uncharged are left in the original pool to be separated. The utility model has the advantages that the separation equipment is simple, the utility model is easy to be operated and grasped, the cost is low, the separation time is short, and the efficiency is high.

Description

Protein concentrates/purification devices

Technical field:

The utility model relates to chemical apparatus, and electric gel bridge isolation methods such as specifically a kind of utilization concentrate/device of protein purification.

Background technology:

At present proteinic concentrating/purification process is more, mainly contain salting-out process, isoelectric point precipitation, low temperature organic solvent precipitation method, dialysis and ultrafiltration, gel filtration method, ion exchange chromatography etc., every kind of method all has the equipment of oneself, the operating process that these equipment have is complicated, the separation and purification time that has is longer, and the purifying cost that has is higher.

The utility model content:

The purpose of this utility model is for overcoming the deficiencies in the prior art, and provides a kind of protein to concentrate/purification devices, and this apparatus structure is simple, and is easy to operate, and sepn process is fast, the purifying cost is low.

The utility model mainly by direct supply, wait to concentrate/the former pond of protein purification, anodal pond, negative pole pond and gel glass bridge tube form.The positive and negative electrode of anodal pond, negative pole pond and direct supply is connected.Anodal pond, negative pole pond respectively by the gel glass bridge tube with wait to concentrate/the former pond of protein purification solution is communicated with; All can under on anodal pond, the negative pole pond in former pond.

Described gel glass bridge tube is sealed its lower end with 1% agar, condensation 30 minutes, and the squeegee of filling 1～12% is poured into 4～15cm, and water is sealed, and condensation was poured out water in 30 minutes, blotted, and condensation is standby after 30 minutes.During use, the glass bridge tube will be immersed in positive and negative electrode pond and the former pond.

Damping fluid 5ml is poured in described anodal pond, negative pole pond into, and former pond adds damping fluid 100～150ml, again the protein soln of purifying to be separated is added former pond.

Damping fluid is a pH value 2～9 in the described pond: by 36.6gTris, the HCl of 48ml 1mol/l or add water to 80ml earlier transfers PH with dense HCl, is settled to 100ml again and makes.

Described squeegee is by 4.9ml distilled water, the acrylamide of 1～6.0ml30%, and 3.8ml1.5mol/lTris-HCl, the ammonium persulphate of 150 μ l 10%, 6 μ l TEMED are formulated.

Described volts DS is 50～300V.

After the protein purification instrument installs, in 4 ℃ of environment, add electrophoretic voltage, this moment, protein and amino acid to be separated entered anodal pond and negative pole pond by the character of its electric charge from former pond by the gel bridge tube respectively, and iso-electric point equals the protein of former pond PH and then stays Yuan Chi.By regulating the pH value in former pond, make the protein of different iso-electric points under electric field action, enter the positive and negative electrode pond, till purifying.

The standard of separation and purification: it is standard that lipidated protein single district band occurs with SDS-acrylamide gel electrophoresis collection of illustrative plates; It is standard that amino acid purity occurs unimodal with amino acidanalyser.

Advantage of the present utility model is: the concentrating and purifying process novelty, and processing ease, separating device is simple, and cost is low, the shortening of separation and purification time, efficient height.

Description of drawings:

Fig. 1 is the utility model structural representation.

1 is that dc-battery, 2 is that anodal pond, 3 is that negative pole pond, 4 is that gel glass bridge tube, 5 is the former pond of purifying substance to be separated.

Embodiment:

The utility model by direct supply 1, wait to concentrate/the former pond 5 of purifying substance, anodal pond 2, negative pole pond 3 and gel glass bridge tube 4 form, anodal pond 2, negative pole pond 3 are connected with the positive and negative electrode of direct supply 1, anodal pond 2, negative pole pond 3 respectively by gel glass bridge tube 4 with wait to concentrate/the former pond 5 of purification solution is communicated with.

Wherein direct supply voltage is 100V, and former pond 5 is 1000 centimetres ³, anodal pond 2, negative pole pond 3 are 1 centimetre ³The gel glass bridge tube is sealed its lower end with 1% agar, condensation 30 minutes, and the squeegee of filling 1～12% is poured into 4～15cm, and water is sealed, and condensation was poured out water in 30 minutes, blotted, and condensation is standby after 30 minutes.During use, the glass bridge tube will be immersed in positive and negative electrode pond and the former pond; Damping fluid 5ml is poured in anodal pond, negative pole pond into, and former pond adds damping fluid 100～150ml, again the protein soln of purifying to be separated is added former pond; Damping fluid in the pond (pH value 2～9): by 36.6gTris, the HCl of 48ml 1mol/l or add water to 80ml earlier transfers PH with dense HCl, is settled to 100ml again and makes; Squeegee is by 4.9ml distilled water, the acrylamide of 1～6.0ml30%, and 3.8ml1.5mol/l Tris-HCl, the ammonium persulphate of 150 μ l 10%, 6 μ l TEMED are formulated.

Claims

1. protein concentrates/purification devices, it is characterized in that: it mainly by direct supply (1), anodal pond (2), negative pole pond (3), etc. the former pond of protein (5) of electric gel bridge tube (4), purifying to be separated form, the positive and negative electrode of direct supply (1) is connected with anodal pond (2), negative pole pond (3) respectively, and gel bridge tube (4) is communicated with anodal pond (2) respectively and is immersed in anodal pond (2), negative pole pond (3), former pond (5) with former pond (5), gel bridge tube (4) with former pond (5), negative pole pond (3).

2. protein according to claim 1 concentrates/purification devices, and it is characterized in that: the former pond of the protein of described purifying to be separated (5), anodal pond (2), negative pole pond (3), its inside is the splendid attire damping fluid all.