Preparation method of protein blot membrane regeneration liquor

A regeneration solution and imprinting technology, which is applied in the field of preparation of protein imprinting membrane regeneration solution, can solve the problems of high antigen elution intensity, harm to operators, weakened protein signal, etc. Effect

Inactive Publication Date: 2010-09-15
TIANGEN BIOTECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The protein imprinting membrane regeneration method introduced in traditional molecular cloning is too strong for antigen elution, which often leads to the weakening of protein signals transferred to solid phase supports such as nitrocellulose

Method used

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Examples

Experimental program
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Effect test

Example Embodiment

[0015] The preparation method of the protein imprinting membrane regenerating solution provided by the present invention includes the following steps:

[0016] (1) Weigh glycine and sodium dodecyl sulfonate separately, and prepare them into an aqueous solution so that the molar concentration of glycine is 25 millimoles / liter and the mass percentage concentration of sodium dodecyl sulfonate is 1%;

[0017] (2) Adjust the pH value of the above aqueous solution to 2 with hydrochloric acid.

Example Embodiment

[0019] Example 1:

[0020] Method of preparing 1 liter of protein imprinting membrane regeneration solution:

[0021] (1) Weigh 1.5 grams of glycine and 10 grams of sodium dodecyl sulfonate respectively;

[0022] (2) Add a certain volume of ultrapure water to completely dissolve;

[0023] (3) Adjust the pH value to 2 with hydrochloric acid;

[0024] (4) Add ultrapure water to make the volume to 1 liter.

Example Embodiment

[0025] Example 2:

[0026] Method of preparing 10 liters of protein imprinting membrane regeneration solution:

[0027] (1) Use a balance to weigh 15 grams of glycine and 100 grams of sodium dodecyl sulfonate;

[0028] (2) Add a certain volume of ultrapure water to completely dissolve;

[0029] (3) Adjust the pH value to 2 with hydrochloric acid;

[0030] (4) Add ultrapure water to make the volume to 10 liters.

[0031] The following introduces the use method of the protein imprinting membrane regenerating solution prepared by the method of the present invention:

[0032] 1. Fully immerse the nitrocellulose membrane or polyvinylidene fluoride membrane detected by the chemiluminescence method in the protein imprinting experiment in the protein imprinting membrane regeneration solution prepared by the method of the present invention, and incubate at room temperature for 15-30 minutes and shake.

[0033] Note: Optimizing the incubation time and temperature can get better results. Generally, h...

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Abstract

The invention relates to a preparation method of protein blot membrane regeneration liquor, belonging to the technical field of biological detection. The preparation method comprises the following steps of: respectively weighing glycine and sodium dodecyl sulfate and preparing into an aqueous solution by ensuring that the molar concentration of the glycine is 20 millimole/liter, and the mass percent concentration of the sodium dodecyl sulfate is 1 percent; and regulating the pH of the aqueous solution by using hydrochloric acid. The protein blot membrane regeneration liquor prepared by using the method has strong efficacy and can efficiently and specifically dissociate the combination of antigens and antibodies but has moderate action on protein combined onto the membrane. The regeneration liquor does not contain beta-mercaptoethanol, has no poison and smell, no toxic hazard to experiment operators, simple use method, quick operation and convenient storage and can be stored at room temperature.

Description

technical field [0001] The invention relates to a method for preparing protein imprinted membrane regeneration solution, belonging to the technical field of biological detection. Background technique [0002] Western blotting is an experimental method commonly used in molecular biology, biochemistry and immunogenetics. Its basic principle is to stain the cells or biological tissue samples treated by gel electrophoresis through specific antibodies, and analyze the coloring position and Depth of Staining Obtain information on how specific proteins are expressed in the cells or tissues being analyzed. The specific test method is to transfer the protein sample separated by polyacrylamide gel electrophoresis to a solid phase support (such as nitrocellulose membrane or polyvinylidene fluoride membrane). Keep the types of peptides separated by electrophoresis and their biological activities unchanged. Use the protein or polypeptide on the solid phase carrier as the antigen, react...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/76
Inventor 龙晶俞萍李晓晨孙克非
Owner TIANGEN BIOTECH BEIJING
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