CN112444552B - Color-changeable polyacrylamide gel and kit containing same - Google Patents

Abstract

The invention discloses a color-changeable polyacrylamide gel and a kit containing the same, wherein the color-changeable polyacrylamide gel comprises separation glue and concentrated glue, wherein the separation glue comprises 6 to 15 percent of polyacrylamide, 0.375M Tris-HCl, 0.1 percent of SDS, 10 percent of ammonium persulfate, 0.1 percent of tetraethylethylenediamine and 0.002 to 0.01 percent of colorant; the concentrated gel comprises 5% polyacrylamide, 0.25M Tris-HCl, 0.1% SDS, 0.1% ammonium persulfate, 0.1% tetraethylethylenediamine and 0.02% to 0.1% colorant. The design of reducing the toxicity of the accelerator in the preparation process of the color-changeable polyacrylamide gel ensures that experimenters are not damaged by volatile toxic compounds; meanwhile, the coloring agents are respectively added into the separation gel and the concentrated gel, so that a user can conveniently judge whether the concentrated gel and the separation gel are solidified and loaded, the protein separation effect is not influenced, the gel preparation reagents are mixed in advance, the gel preparation process is accelerated, and the experiment efficiency is improved.

Description

Color-changeable polyacrylamide gel and kit containing same

Technical Field

The invention belongs to the technical field of electrophoresis, and particularly relates to a color-changeable polyacrylamide gel kit.

Background

Polyacrylamide gel electrophoresis (PAGE) is the most extensive and fundamental experiment among the current protein experiments, and includes Native-PAGE and SDS-PAGE. According to the traditional PAGE, acrylamide and methylene acrylamide are polymerized by taking Tetramethylethylenediamine (TEMED) as an accelerator under the catalytic action of Ammonium Persulfate (APS), in the polymerization process, TEMED catalyzes ammonium persulfate to generate free radicals which initiate polymerization of acrylamide monomers, and methylene bond crosslinking is generated between methylene bisacrylamide and acrylamide chains to form a three-dimensional network structure, so that the acrylamide gel is prepared.

The traditional acrylamide gel has the following defects that TEMED is volatile, inflammable, corrosive, and strong in neurotoxicity and pungent smell; the concentrated gel is colorless, so that a sample is easily added out of a sample hole during sample loading, thereby causing pollution among samples and influencing the identification result after protein separation; whether the separation glue and the concentrated glue are solidified cannot be judged, the time is generally waited for 30min to 1h, and a large amount of time is wasted; if the waiting time is reduced, the gel preparation is easy to fail because the separation gel and the concentrated gel are not completely solidified; the glue making process is complex, various solutions need to be mixed, the glue making process is time-consuming and labor-consuming, and the experiment efficiency is reduced. In conclusion, a safer and more efficient rapid polyacrylamide gel preparation kit is urgently needed for the traditional polyacrylamide gel.

Disclosure of Invention

The purpose of the invention is as follows: in order to solve the technical problems in the prior art, the invention provides a color-changeable polyacrylamide gel and a kit containing the same.

In order to achieve the purpose, the technical scheme of the invention is as follows:

a colored polyacrylamide gel comprising a separation gel and a concentration gel, wherein the separation gel comprises 6-15% polyacrylamide, 0.375M Tris-HCl, 0.1% sds, 10% ammonium persulfate, 0.1% tetraethylethylenediamine, 0.002% to 0.01% colorant; the concentrated gum comprises 5% polyacrylamide, 0.25M Tris-HCl, 0.1% SDS, 0.1% ammonium persulfate, 0.1% tetraethylethylenediamine and 0.02% to 0.1% colorant.

Specifically, the polyacrylamide is polymerized from acrylamide and methylene bisacrylamide according to a mass ratio of 29.

The pH value of the separation gel is 8.8, and the pH value of the concentration gel is 6.8.

Specifically, the colorant is processed Direct red 80, wherein the Direct red 80 is processed as follows:

(1) Taking powder Direct red 80, dissolving the powder with ultrapure water, and centrifuging to obtain an upper layer solution;

(2) Sequentially carrying out activated carbon adsorption and gelatin adsorption, and centrifuging to obtain a supernatant solution;

(3) And (4) freeze-drying the supernatant to obtain a solid which is the finally required colorant.

The invention also provides a preparation method of the colored polyacrylamide gel, which comprises the following steps:

(1) Preparing a stock solution: the stock solution included 30% acr-Bis, 4 × upper gel buffer, 4 × lower gel buffer and ammonium persulfate solution;

(2) Preparing polyacrylamide separation gel: mixing Acr-Bis 30% of formula amount, 0.1% SDS, 0.375M Tris-HCl, 0.1% tetraethylethylenediamine and 0.002-0.01% coloring agent, adding ammonium persulfate, mixing, injecting into the gap between two glass plates, and adding alcohol or water to cover the separating gel;

(3) Preparing polyacrylamide concentrated gel: taking 30 percent of Acr-Bis, 0.25M Tris-HCl, 0.1 percent of SDS, 0.1 percent of tetraethylethylenediamine and 0.02 to 0.1 percent of coloring agent according to the formula amount, fully mixing, adding ammonium persulfate, fully mixing, pouring out the covering layer liquid after the separation gel is completely polymerized, washing by deionized water, filling the prepared concentration gel on the separation gel, immediately inserting a clean matched comb to avoid bubbles, and pulling out the comb after the concentration gel is polymerized to form a sample adding hole.

The invention further provides a color polyacrylamide gel kit, which consists of 30% of Acr-Bis, 4 x upper layer gel buffer solution, 4 x lower layer gel buffer solution and ammonium persulfate, wherein the 4 x upper layer gel buffer solution consists of 0.25M Tris-HCl, 0.1% SDS, 0.1% tetraethylethylenediamine and 0.02% -0.1% of colorant, and the 4 x lower layer gel buffer solution consists of 0.375M Tris-HCl, 0.1% SDS, 0.1% tetraethylethylenediamine and 0.002% -0.01% of colorant.

Wherein the colorant is processed Direct red 80, wherein the Direct red 80 is processed as follows:

(1) Dissolving powder Direct red 80 in ultrapure water, and centrifuging to obtain an upper layer solution;

(2) Sequentially carrying out activated carbon adsorption and gelatin adsorption, and centrifuging to obtain a supernatant solution;

(3) And (4) freeze-drying the supernatant to obtain a solid which is the finally required colorant.

Has the advantages that: compared with the prior art, the invention has the following advantages:

(1) The accelerator is premixed in the 4 multiplied upper layer glue buffer solution and the 4 multiplied lower layer glue buffer solution, so that the design of reducing the toxicity of the accelerator ensures that experimenters are not damaged by volatile toxic compounds;

(2) The colorant is directly added into the 4 multiplied upper layer gel buffer solution and the 4 multiplied lower layer gel buffer solution, so that a user can conveniently judge whether the concentrated gel and the separation gel are solidified and loaded, and the protein separation effect is not influenced;

(3) And the glue preparation reagents are mixed in advance, so that the glue preparation process is accelerated, and the experiment efficiency is improved.

Drawings

FIG. 1 is a 10% colored polyacrylamide gel prepared in example 3, with the upper layer being overlying alcohol or water and the lower layer being a light red separating gel (not solidified);

FIG. 2A is a 10% colored polyacrylamide gel prepared in example 3, with the upper layer being a red concentrated gel (uncured) and the lower layer being a colorless isolated gel (cured);

FIG. 2B is a 10% colored polyacrylamide gel prepared in example 3, with the upper layer being a pink concentrated gel (set) and the lower layer being a colorless isolated gel (set);

FIG. 3 is a 10% color polyacrylamide gel prepared in example 3 after electrophoresis with a Marker added thereto and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample;

FIG. 4 is a graph of a 10% polyacrylamide gel prepared in comparative example 1 after electrophoresis with a Marker added thereto and a protein sample, lane 1 being the Marker, lane 2 being the protein sample;

FIG. 5 is a graph of 6% colored polyacrylamide gel prepared in example 1 after electrophoresis with Marker and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample;

FIG. 6 is a graph of an 8% colored polyacrylamide gel prepared in example 2 after electrophoresis with a Marker added thereto and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample;

FIG. 7 is a drawing of the 12% colored polyacrylamide gel prepared in example 4 after electrophoresis with a Marker attached to the gel and a protein sample, wherein Marker is shown in lane 1 and the protein sample is shown in lane 2;

FIG. 8 is a 15% colored polyacrylamide gel prepared in example 5 after electrophoresis with a Marker attached thereto and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample.

Detailed Description

Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The experimental procedures, in which specific conditions are not specified in the examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.

The Marker used in the examples is a pre-dyed color protein molecular weight Marker of Kyowa Kayji Biotechnology GmbH, cat # KGM471; the protein sample was Bovine Serum Albumin (BSA).

According to different optimal separation ranges required by different protein samples, experimenters generally select gels with different concentrations to meet the experimental requirements, and the invention can prepare the gels within the concentration range of 6-15%.

Example 1% colored polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis: taking 29 g of acrylamide and 1 g of methylene bisacrylamide for 100ml, adding water to dissolve, metering the volume to 100ml, and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.4% SDS, 0.1% tetraethylethylenediamine and 0.02-0.1% colorant, pH 6.8;

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% SDS, 0.4% tetraethylethylenediamine and 0.002% to 0.01% colorant, pH 8.8;

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

Preparation method of 2.6% color polyacrylamide separation gel

2.7ml of distilled water, 30% Acr-Bis 1.0ml of the mixture, and 1.25ml of 4 Xlower layer gel buffer solution were mixed well, and then 50ul of 10% ammonium persulfate was added thereto, and mixed well, and then injected into the gap between the two glass plates, and water was added to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Mixing distilled water 1.17ml and 30% Acr-Bis 0.33ml and 4 × supernatant buffer 0.5ml, adding 10% ammonium persulfate 0.02ml, mixing, pouring out the covering layer liquid after the gel turns from light red to colorless, pouring the prepared concentrated gel on the gel, immediately inserting clean matched tree to avoid bubbles, and after the gel turns from red to pink, pulling out a comb to form a sample hole, thereby performing SDS-PAGE electrophoresis experiment.

FIG. 5 is a 6% color polyacrylamide gel prepared in example 1 after electrophoresis with a Marker added thereto and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample.

Example 2% color polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis:100ml, taking 29 g of acrylamide and 1 g of methylene bisacrylamide, adding water to dissolve, and fixing the volume to 100ml and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.4% SDS, 0.1% tetraethylethylenediamine and 0.02-0.1% colorant, pH 6.8;

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% SDS, 0.4% tetraethylethylenediamine and 0.002-0.01% colorant, pH 8.8;

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

2. Preparation method of 8% colored polyacrylamide separation gel

2.4ml of distilled water, 30 percent of Acr-Bis 1.3ml and 1.25ml of 4 multiplied lower layer gel buffer solution are taken, fully and evenly mixed, 50ul of 10 percent ammonium persulfate is added, fully and evenly mixed and rapidly injected into the gap between the two glass plates, and then water is added to cover the separation gel.

3. Preparation method of 5% polyacrylamide concentrated gel

Taking 1.17ml of distilled water, mixing 30% of Acr-Bis 0.33ml and 4 multiplied by upper layer glue buffer solution 0.5ml uniformly, adding 0.02ml of 10% ammonium persulfate, mixing uniformly, pouring out the covering layer liquid after the separation glue is changed from light red to colorless, pouring the prepared concentration glue on the separation glue, immediately inserting clean matched trees to avoid bubbles, and pulling out a comb after the concentration glue is changed from red to pink to form a sample adding hole, thus carrying out SDS-PAGE electrophoresis experiment.

FIG. 6 is a graph of the 8% colored polyacrylamide gel prepared in example 2 after electrophoresis with Marker added to the protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample.

Example 3 configuration of 10% colored polyacrylamide gel.

1. Each mixed stock solution component:

1) 30% Acr-Bis: taking 29 g of acrylamide and 1 g of methylene bisacrylamide for 100ml, adding water to dissolve, metering the volume to 100ml, and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.4% SDS, 0.1% tetraethylethylenediamine and 0.02-0.1% colorant, pH 6.8

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% SDS, 0.4% tetraethylethylenediamine and 0.002-0.01% colorant, pH 8.8

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

Preparation method of 2.10% color polyacrylamide separation gel

2.0ml of distilled water was taken, 30% of Acr-Bis 1.7ml and 4X lower layer gel buffer 1.25ml were mixed well, 50ul of 10% ammonium persulfate was added and mixed well, and injected into the gap between the two glass plates quickly, and water was added to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Mixing distilled water 1.17ml,30% Acr-Bis 0.33ml,4 × supernatant buffer 0.5ml, adding 10% ammonium persulfate 0.02ml, mixing, pouring out the covering layer liquid when the gel turns from light red to colorless, pouring the prepared concentrated gel on the gel, immediately inserting clean matched tree to avoid bubbles, and pulling out a comb to form a sample hole, thereby performing SDS-PAGE electrophoresis experiment.

FIG. 1 is a 10% colored polyacrylamide gel prepared according to example 6, with the upper layer being overlying alcohol or water and the lower layer being a light red colored release gum (not solidified);

FIG. 2A is a 10% colored polyacrylamide gel prepared in example 6, with the upper layer being a red concentrated gel (uncured) and the lower layer being a colorless isolated gel (cured);

FIG. 2B shows a 10% colored polyacrylamide gel prepared in example 6, with a red concentrated gel (solidified) on the top layer and a colorless isolated gel (solidified) on the bottom layer.

Thus, the color change of the separation gel and the concentration gel can be directly used to judge whether the gel is solidified and when the sample should be added.

Example 4% colored polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis: taking 29 g of acrylamide and 1 g of methylene bisacrylamide for 100ml, adding water to dissolve, metering the volume to 100ml, and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.4% SDS, 0.1% tetraethylethylenediamine and 0.02-0.1% colorant, pH 6.8;

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% SDS, 0.4% tetraethylethylenediamine and 0.002-0.01% colorant, pH 8.8;

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

Preparation method of 2.12% colored polyacrylamide separation gel

1.7ml of distilled water, 30% Acr-Bis 2.0ml, and 1.25ml of 4 Xlower layer gel buffer were mixed well, then 50ul of 10% ammonium persulfate was added and mixed well, and injected into the gap between the two glass plates quickly, and water was added to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Taking 1.17ml of distilled water, mixing 30% of Acr-Bis 0.33ml and 4 multiplied by upper layer glue buffer solution 0.5ml uniformly, adding 0.02ml of 10% ammonium persulfate, mixing uniformly, pouring out the covering layer liquid after the separation glue turns into colorless with light red, then pouring the prepared concentration glue on the separation glue, immediately inserting clean matched trees to avoid bubbles, and after the concentration glue changes from red to pink, pulling out a comb to form a sample adding hole, thus carrying out SDS-PAGE electrophoresis experiment.

FIG. 7 is a 12% color polyacrylamide gel prepared in example 4 after electrophoresis with Marker added and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample.

Example 5% colored polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis:100ml, taking 29 g of acrylamide and 1 g of methylene bisacrylamide, adding water to dissolve, and fixing the volume to 100ml and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.4% SDS, 0.1% tetraethylethylenediamine and 0.02-0.1% colorant, pH 6.8

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% SDS, 0.4% tetraethylethylenediamine and 0.002-0.01% colorant, pH 8.8

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

Preparation method of 2.15% colored polyacrylamide separation gel

Mixing distilled water 1.2ml,30% Acr-Bis 2.5ml,4 × lower layer gel buffer 1.25ml, adding 10% ammonium persulfate 50ul, mixing, injecting into the gap between two glass plates, and adding water to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Taking 1.17ml of distilled water, mixing 0.33ml of Acr-Bis 0.33ml of distilled water and 0.5ml of 4 multiplied upper layer gel buffer solution fully, adding 0.02ml of 10 percent ammonium persulfate, mixing fully, pouring out the covering layer liquid after the separation gel is changed from light red to colorless, pouring the prepared concentration gel on the separation gel, immediately inserting clean matched trees to avoid bubbles, and pulling out a comb to form a sample adding hole after the concentration gel is changed from red to pink, thus carrying out SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) experiment.

FIG. 8 is a 15% colored polyacrylamide gel prepared in example 5 after electrophoresis with a Marker attached thereto and a protein sample, wherein lane 1 is the Marker and lane 2 is the protein sample.

Example 6% color native polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis:100ml, taking 29 g of acrylamide and 1 g of methylene bisacrylamide, adding water to dissolve, and fixing the volume to 100ml and mixing uniformly;

2) 4 × supernatant buffer: 30ml,1M Tris-HCl, 0.1% tetraethyl ethylenediamine and 0.02-0.1% colorant, pH 6.8

3) 4 × lower layer gel buffer: 30ml,1.5M Tris-HCl, 0.4% tetraethylethylenediamine and 0.002-0.01% coloring agent, pH 8.8

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is removed, 1ml of water is added for dissolving and mixing evenly.

Preparation method of 2.10% color non-denatured polyacrylamide separation gel

2.0ml of distilled water was taken, 30% of Acr-Bis 1.7ml and 4X lower layer gel buffer 1.25ml were mixed well, 50ul of 10% ammonium persulfate was added and mixed well, and injected into the gap between the two glass plates quickly, and water was added to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Taking 1.17ml of distilled water, mixing 0.33ml of Acr-Bis 0.33ml of distilled water and 0.5ml of 4 multiplied upper layer gel buffer solution fully, adding 0.02ml of 10 percent ammonium persulfate, mixing fully, pouring out the covering layer liquid after the separation gel is changed from light red to colorless, pouring the prepared concentration gel on the separation gel, immediately inserting clean matched trees to avoid bubbles, and pulling out a comb after the concentration gel is changed from red to pink to form a sample adding hole, thus carrying out the next polyacrylamide gel electrophoresis experiment.

The concentration of acrylamide and methylene acrylamide in the solution is determined based on other concentrations of 6-15%, so that the non-denaturing polyacrylamide gel in the concentration range can be realized according to the formula of the invention, and the specific steps are the same.

Comparative example 1% polyacrylamide gel formulation.

1. Each mixed stock solution component:

1) 30% Acr-Bis:100ml, taking 29.2 g of acrylamide and 0.8 g of methylene bisacrylamide, adding water to dissolve, and fixing the volume to 100ml and mixing uniformly;

2) 2MTris-Hcl buffer: 100ml, taking 24.2228 g of Tris, adding a proper amount of water for dissolution, adjusting the pH value to 8.8 by using Hcl, fixing the volume to 100ml, and uniformly mixing.

3) 1M Tris-HCl buffer: 50ml, taking 6.057 g of Tris, adding a proper amount of water for dissolution, adjusting the pH value to 6.8 by using Hcl, metering the volume to 50ml, and uniformly mixing.

4) 10% SDS:5ml, 0.5 g of SDS is taken, dissolved by adding water and the volume is adjusted to 5ml.

4) Ammonium persulfate: 10 percent, 0.1 g of ammonium persulfate is taken and dissolved in 1ml of water to be mixed evenly.

Preparation method of 2.10% polyacrylamide separation gel

2.2ml of distilled water was taken, 30% of Acr-Bis 1.7ml,2M Tris-HCl buffer 1ml,10% of SDS 50ul was thoroughly mixed, 10% of ammonium persulfate 50ul was added, and thoroughly mixed, 2ul of TEMED was added and thoroughly mixed, and then the mixture was rapidly poured into the space between the two glass plates, and further alcohol or water was added to cover the separation gel.

Preparation method of 3.5% polyacrylamide concentrated gel

Taking 1.4ml of distilled water, 30 percent of Acr-Bis 0.33ml,1M Tris-HCl buffer solution 250ul,10 percent of SDS 50ul, fully mixing, adding 20ul of 10 percent ammonium persulfate, fully mixing, adding 2ul of TEMED to completely polymerize the separation gel, pouring out covering layer liquid, pouring the prepared concentration gel on the separation gel, immediately inserting clean matched trees to avoid bubbles, pulling out a comb after the concentration gel is polymerized, forming sample adding holes, and carrying out the next polyacrylamide gel electrophoresis experiment.

FIG. 4 is a graph of a 10% polyacrylamide gel prepared in comparative example 1 after electrophoresis with a Marker added thereto and a protein sample, wherein Marker is in lane 1 and the protein sample is in lane 2. From the separation effect of the Marker and the protein sample shown in fig. 3 and 4, the polyacrylamide gel prepared by the present invention has substantially the same separation effect as the conventional polyacrylamide gel.

The separation gel disclosed by the invention contains 0.002-0.01% of coloring agent, so that the color of the separation gel is changed before and after solidification, the solidification degree of the separation gel is conveniently judged, the concentrated gel is conveniently prepared in time, and the separation effect of protein is not influenced; the concentrated glue contains 0.02-0.1% of colorant, so that the color of the concentrated glue changes before and after solidification, the solidification degree of the concentrated glue can be conveniently judged, and a user can conveniently sample the concentrated glue without influencing the separation effect of the protein.

The present invention provides a method for preparing colored polyacrylamide gel, and a plurality of methods and ways for implementing the technical scheme, and the above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention. All the components not specified in the present embodiment can be realized by the prior art.

Claims (5)

1. The variable-color polyacrylamide gel is characterized by comprising separation gel and concentration gel, wherein the separation gel comprises 6-15% of polyacrylamide, 0.375M Tris-HCl, 0.1% of SDS, 0.1% of ammonium persulfate, 0.1% of tetraethylethylenediamine and 0.002% -0.01% of colorant; the concentrated gel comprises 5% of polyacrylamide, 0.25M Tris-HCl, 0.1% of SDS, 0.1% of ammonium persulfate, 0.1% of tetraethylethylenediamine and 0.02% -0.1% of coloring agent, wherein the coloring agent is treated Direct red 80, and the Direct red 80 is treated as follows:

(1) Dissolving powder Direct red 80 in ultrapure water, and centrifuging to obtain an upper layer solution;

(2) Sequentially carrying out activated carbon adsorption and gelatin adsorption, and centrifuging to obtain a supernatant solution;

(3) And (4) freeze-drying the supernatant to obtain a solid which is the finally required colorant.

2. The color-changeable polyacrylamide gel according to claim 1 wherein the polyacrylamide is polymerized from acrylamide and methylene bisacrylamide in a mass ratio of 29.

3. The color-changeable polyacrylamide gel of claim 1 wherein the pH of said separation gel is 8.8 and the pH of said concentration gel is 6.8.

4. A method for preparing a variable color polyacrylamide gel according to any one of claims 1-3, comprising the steps of:

(1) Preparing a stock solution: the stock solution comprises 30% of Acr-Bis, 4 multiplied by upper layer glue buffer solution, 4 multiplied by lower layer glue buffer solution and ammonium persulfate solution;

(2) Preparing polyacrylamide separation gel: taking 30 percent of Acr-Bis, 0.1 percent of SDS, 0.375M of Tris-HCl, 0.1 percent of tetraethyl ethylenediamine and 0.002 to 0.01 percent of colorant according to the formula amount, fully mixing, adding ammonium persulfate, fully mixing, then quickly injecting into a gap between two glass plates, and then adding alcohol or water to cover on separation gel;

(3) Preparing polyacrylamide concentrated gel: taking 30% of Acr-Bis, 0.25M Tris-HCl, 0.1% of SDS, 0.1% of tetraethylethylenediamine and 0.02% -0.1% of coloring agent according to the formula amount, fully and uniformly mixing, adding ammonium persulfate, fully and uniformly mixing, pouring out the covering layer liquid after the separation gel is completely polymerized, washing with deionized water, pouring the prepared concentration gel on the separation gel, immediately inserting a clean matched comb to avoid bubbles, and pulling out the comb after the concentration gel is polymerized to form a sample adding hole.

5. The variable-color polyacrylamide gel kit is characterized by comprising 30% of Acr-Bis, 4 multiplied by upper gel buffer solution, 4 multiplied by lower gel buffer solution and ammonium persulfate, wherein the 4 multiplied by upper gel buffer solution comprises 0.25M Tris-HCl, 0.1% SDS, 0.1% tetraethylethylenediamine and 0.02% -0.1% of colorant, and the 4 multiplied by lower gel buffer solution comprises 0.375M Tris-HCl, 0.1% SDS, 0.1% tetraethylethylenediamine and 0.002% -0.01% of colorant; the colorant is processed Direct red 80, wherein the Direct red 80 is processed as follows:

(1) Taking powder Direct red 80, dissolving the powder with ultrapure water, and centrifuging to obtain an upper layer solution;

(2) Sequentially carrying out activated carbon adsorption and gelatin adsorption, and centrifuging to obtain a supernatant solution;

(3) And (4) freeze-drying the supernatant to obtain a solid which is the finally required colorant.

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