Polyacrylamide gel and kit thereof

A technology of polyacrylamide gel and polyacrylamide, which is applied in the direction of measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of electrophoresis speed drop, unstable solution performance, sample contamination, etc., and achieve stable gel Solidification speed, clear protein bands, and convenient placement at room temperature

Active Publication Date: 2018-06-19
CHONGQING KANGZHI PHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 1. TEMED is volatile, flammable, corrosive, and has strong neurotoxicity and irritating odor
[0004] 2. The preparation process is complicated, and various solutions need to be mixed
The glue making process is time-consuming, which greatly reduces the experimental efficiency of the experimenters, and the strong pungent smell of TEMED added later will have a bad impact on the health of the experimenters
[0005] 3. The gel speed is slow. The gel speed of traditional separating gel is 30min-1h, and that of stacking gel is about 30min.
[0006] 4. The stacking gel is colorless, and it is easy to add the sample to the outside of the sample hole

Method used

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  • Polyacrylamide gel and kit thereof
  • Polyacrylamide gel and kit thereof
  • Polyacrylamide gel and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 6% polyacrylamide gel preparation

[0041] 1. Components of each mixed stock solution:

[0042] 1) Separating gel solution A: 100ml, take 11.6 grams of acrylamide, 0.4 grams of methylenebisacrylamide, add water and stir to dissolve, then dilute to 100ml and mix well;

[0043] 2) Separating gel solution B: 100ml, wherein 0.75MTris, 0.2wt% SDS, 0.06wt% tetraethylethylenediamine, 0.3wt% sodium bisulfite, pH value is 8.8.

[0044] 3) Concentrating gel solution A: 20ml, take 2 grams of acrylamide, 0.0533 grams of methylenebisacrylamide, add water, stir to dissolve, dilute to 20ml and mix well;

[0045] 4) Stacking gel solution B: 20ml of which 0.25MTris, 0.2wt% SDS, 0.1wt% sodium bisulfite, 0.2wt% tetraethylethylenediamine, pH value is 6.8.

[0046] 5) Ammonium persulfate (AP): 10 wt%, take 0.1 g of ammonium persulfate, add 1 ml of water to dissolve and mix.

[0047] After the preparation of the above reagents, place at 4°C for 10 days before preparing the ge...

Embodiment 28

[0053] Embodiment 2 8% polyacrylamide gel preparation

[0054] 1. Components of each mixed stock solution:

[0055] 1) Separating gel solution A: 50ml, take 8 grams of acrylamide, 0.2133 grams of methylenebisacrylamide, add water and stir to dissolve, then dilute to 50ml and mix well;

[0056] 2) Separating gel solution B: 50ml, wherein 0.7MTris, 0.2wt% SDS, 0.08wt% tetraethylethylenediamine, 0.35wt% sodium bisulfite, pH value is 8.8.

[0057] 3) Concentrating gel solution A: 10ml, take 0.9733g of acrylamide and 0.0267g of methylenebisacrylamide, add water and stir to dissolve, then dilute to 10ml and mix well;

[0058] 4) Stacking gel solution B: 10ml of which 0.3MTris, 0.2wt% SDS, 1wt% sodium bisulfite, 0.2wt% sodium bisulfite, 0.16wt% tetraethylethylenediamine, pH value is 6.8.

[0059] 5) Ammonium persulfate (AP): 10 wt%, take 0.1 g of ammonium persulfate, add 1 ml of water to dissolve and mix.

[0060] After the preparation of the above reagents, place at 4°C for 15 da...

Embodiment 310

[0066] Embodiment 3 10% polyacrylamide gel preparation

[0067] 1. Components of each mixed stock solution:

[0068] 1) Separating gel solution A: 100ml, take 19.4667 grams of acrylamide and 0.5333 grams of methylenebisacrylamide, add water and stir to dissolve, then dilute to 100ml and mix well;

[0069] 2) Separating gel solution B: 100ml, wherein 0.8MTris, 0.2wt% SDS, 0.064wt% tetraethylethylenediamine, 0.25wt% sodium bisulfite, pH value is 8.8.

[0070] 3) Concentrating gel solution A: 20ml, take 1.5573g of acrylamide and 0.04267g of methylenebisacrylamide, add water, stir to dissolve, dilute to 20ml and mix well;

[0071] 4) Concentrating gel solution B: 20ml of which 0.2MTris, 0.2wt% SDS, 0.16wt% tetraethylethylenediamine, 0.1wt% sodium bisulfite, pH 6.8.

[0072] 5) Ammonium persulfate (AP): 10 wt%, take 0.1 g of ammonium persulfate, add 1 ml of water to dissolve and mix.

[0073] After the preparation of the above reagents, place at 4°C for 10 days before preparing ...

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PUM

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Abstract

The invention relates to a polyacrylamide gel. The polyacrylamide gel is prepared from a separation gel and a concentration gel, wherein the separation gel is prepared from 6 to 15wt% of polyacrylamide, 0.35 to 0.4M of tris(hydroxymethyl)methyl aminomethane, 0.03 to 0.2wt% of tetraethylethylenediamine, 0.1 to 1wt% of sodium hydrogen sulfite and 0.08 to 0.15wt% of ammonium persulfate; the concentration gel is prepared from 4 to 5wt% of polyacrylamide, 0.1 to 0.15M of tris(hydroxymethyl)methyl aminomethane, 0.08 to 0.4wt% of tetraethylethylenediamine, 0.05 to 0.5wt% of sodium hydrogen sulfite and 0.08 to 0.15wt% of ammonium persulfate. The polyacrylamide gel has the advantages that under the synergistic function of multiple components, the stable setting speed of the gel is ensured, and thelong-term stability of the gel is improved; the effect of protein separating by gel electrophoresis is good, and the protein stripes are clear; by not adding the volatile poisonous accelerant TEMED, the injury to the experiment person by the volatile poisonous compound is avoided.

Description

technical field [0001] The invention belongs to polyacrylamide gel electrophoresis, in particular to a polyacrylamide gel and a kit thereof. Background technique [0002] Polyacrylamide gel electrophoresis (PAGE) is the most widely used and most basic experiment in biological experiments. It has two forms: non-denaturing polyacrylamide gel electrophoresis (Native-PAGE) and polyacrylamide gel electrophoresis (PAGE). Acrylamide gel (SDS-PAGE). At present, the traditional PAGE is mainly composed of monomer acrylamide and methylene bisacrylamide with ammonium persulfate (APS) as the catalyst and tetramethylethylenediamine (TEMED) as the accelerator. During the polymerization process, TEMED catalyzes Ammonium persulfate generates free radicals, which initiate the polymerization of acrylamide monomers, and at the same time, methylene bisacrylamide and acrylamide chains generate methylene bond crosslinks, thereby forming a three-dimensional network structure and making a gel. How...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 文倩黎波
Owner CHONGQING KANGZHI PHARM
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