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Gradient gel and preparation method thereof

A gel and gradient technology, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve the problems of insufficient transfer, slow electrophoresis speed, hindering the speed of protein molecules, etc., to speed up electrophoresis speed and strengthen wear Permeability, improve protein transfer rate and protein transfer quality

Inactive Publication Date: 2014-02-12
GUANGZHOU WONDFO BIOTECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] However, for the second product, in the process of gel electrophoresis transfer of mixed antigens using western blotting technology, the waiting time for electrophoresis and transfer is longer, and electrophoresis usually takes 4.5 hours, which is time-consuming and less efficient
In addition, if the concentration of the homogeneous gel is set too high, it will hinder the speed of protein molecule electrophoresis and affect the transfer rate; if the concentration is too small, the speed of protein molecule electrophoresis will be fast, and protein molecules of different molecular weights will not be clearly separated during electrophoresis. Open, resulting in protein transfer ineffective, may cause the entire Western blotting process meaningless
At the same time, due to the use of a gel with uniform concentration, whether it is gel electrophoresis or protein transfer, protein molecules of different molecular weights must uniformly pass through the same concentration of gel, the electrophoresis speed is slow, the efficiency is low, and the transfer is not thorough enough. , long waiting time

Method used

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  • Gradient gel and preparation method thereof
  • Gradient gel and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0015] 1. Prepare separation gel: mix H 2 O, acrylamide stock solution, separating gel buffer, 10% SDS, and ammonium persulfate are adjusted to 8%, 10%, 13%, and 15% concentration of separating gel in a certain proportion. The formulations of these two different concentrations of separating gels are as follows:

[0016] (1) The separation gel with a concentration of 8%: 7.8mL H 2 O, 3.3mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL10% SDS, 0.42mL16mL / mL ammonium persulfate;

[0017] (2) The concentration is 15% separating gel: 0.78mL H 2 O, 9.5mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL 10% SDS, 0.42mL 16mL / mL ammonium persulfate.

[0018] 2. Prepare concentrated gel: mix 13.3mL H 2 O, 1.8 mL of acrylamide stacking gel stock solution, 2.3 mL of stacking gel buffer, 0.19 mL of 10% SDS, and 0.42 mL of 16 mL / mL ammonium persulfate were prepared to form a stacking gel.

[0019] 3. Take two test tubes, na...

Embodiment 2

[0021] 1. Prepare separation gel: mix H 2 O, acrylamide stock solution, separating gel buffer, 10% SDS, and ammonium persulfate are adjusted to 8%, 11%, and 15% concentration of separating gel in a certain proportion. The formulations of these three different concentrations of separating gels are as follows:

[0022] (1) The separation gel with a concentration of 8%: 7.8mL H 2 O, 3.3mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL10% SDS, 0.42mL16mL / mL ammonium persulfate;

[0023] (2) The concentration is 11% separating gel: 3.2mL H 2 O, 7.68mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL10% SDS, 0.42mL16mL / mL ammonium persulfate;

[0024] (3) The concentration is 15% separating gel: 0.78mL H 2 O, 9.5mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL 10% SDS, 0.42mL 16mL / mL ammonium persulfate.

[0025] 2. Prepare concentrated gel: mix 13.3mL H 2 O, 1.8 mL of acrylamide sta...

Embodiment 3

[0028] 1. Prepare separation gel: mix H 2 O, acrylamide stock solution, separating gel buffer, 10% SDS, and ammonium persulfate are adjusted to 8%, 10%, 13%, and 15% concentration of separating gel in a certain proportion. The formulations of these four different concentrations of separating gels are as follows:

[0029] (1) The separation gel with a concentration of 8%: 7.8mL H 2 O, 3.3mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL10% SDS, 0.42mL16mL / mL ammonium persulfate;

[0030] (2) The concentration is 10% separating gel: 3.8mL H 2 O, 6.65mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL10% SDS, 0.42mL16mL / mL ammonium persulfate;

[0031] (3) The separation gel with a concentration of 13%: 1.2mL H 2 O, 9.5mL acrylamide separation gel stock solution, 7.01mL separation gel buffer, 0.19mL 10% SDS, 0.42mL 16mL / mL ammonium persulfate.

[0032] (4) The concentration is 15% separating gel: 0.78mL H 2 O, 9....

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Abstract

The invention discloses a gradient gel and a preparation method thereof. The preparation method is characterized by comprising the following steps: respectively blending water, acrylamide separation gel stock solution, separation gel buffer solution, SDS (sodium dodecyl sulfate) and ammonium persulfate into two or more separation gels with concentration of 8-15% according to a certain proportion, quickly pouring the blended two or more separation gels in different concentrations into a glass plate crack in vertical slab electrophoresis from high to low concentration in order, adding water, adding spacer gel after the separation gel is solidified, and waiting the solidification of the spacer gel. According to the method, the electrophoresis speed is increased, the penetrability of protein molecules is enhanced, and the protein transfer rate and the protein transfer quality are improved.

Description

technical field [0001] The invention discloses a gradient gel and a preparation method thereof. Background technique [0002] Autoantibodies are antibodies directed against one's own tissues, organs, cells and cellular components. The growth, development and survival of the human body are maintained by a complete autoimmune tolerance mechanism, and the normal immune response has a protective defense function, that is, it does not react to its own tissues and components. Once the integrity of self-tolerance is destroyed, the body regards its own tissues and components as "foreign bodies", and an autoimmune reaction occurs, producing autoantibodies. Under certain circumstances, the immune system fails to recognize the body's own substances and regards them as foreign invaders, thereby producing antibodies against these substances (ie, autoantibodies), triggering autoimmunity. These autoantibodies will attack the body's own cells, tissues, and organs, causing inflammatory rea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 王继华
Owner GUANGZHOU WONDFO BIOTECH
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