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In-situ reagent for detection of proteins

A technology of protein and kit, applied in the field of stable protein and/or amino acid detection composition

Inactive Publication Date: 2013-07-10
QUEEN MARY UNIV OF LONDON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The above-mentioned methods all have many drawbacks, such as different protein and / or amino acid specificities, sensitivities, and stability of reagents and / or formed products

Method used

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  • In-situ reagent for detection of proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0148] Embodiment 1: Preparation of o-phthalaldehyde / acetylcysteine ​​reagent

[0149] Dissolve 9.5 g of anhydrous disodium tetraborate in 500 ml of deionized water, add a few drops of 1M NaOH to adjust the pH, and prepare a sodium borate buffer solution with a pH of 9.23. Add 60 mg of N-acetylcysteine ​​into the buffer solution and stir until completely dissolved with a magnetic stirring bar. 60 mg of o-phthalaldehyde was dissolved in 5 ml of methanol, and all of it was added to the above-mentioned sodium borate buffer solution to prepare an acetylcysteine ​​solution. The o-phthalaldehyde / acetylcysteine ​​solution was prepared fresh weekly and refrigerated at 4°C in the dark for 24 hours before use to reduce background fluorescence interference.

Embodiment 2

[0150] Example 2: Preparation of stable o-phthalaldehyde / acetylcysteine ​​for protein detection

[0151] Dissolve 19 g of anhydrous disodium tetraborate in 900 ml of deionized water, add a few drops of 1M NaOH to adjust the pH, and prepare a sodium borate buffer solution with a pH of 9.23. Add 1 ml of TritonX-100, 1.63 g of N-acetylcysteine, and 0.077 g of dithiothreitol into the buffer solution, and stir with a magnetic stirring bar until completely dissolved. 0.268 g of acetylcysteine ​​was dissolved in 10 ml of methanol, and all of it was added to the above-mentioned sodium borate buffer solution to prepare an OPA solution. The solution was made up to 1 L. The o-phthalaldehyde / acetylcysteine ​​solution was refrigerated at 4°C in the dark for 24 hours before use to reduce background fluorescence interference.

Embodiment 3

[0152] Example 3: Preparation of stable o-phthalaldehyde / acetylcysteine, further comprising sodium edetate

[0153] After the same o-phthalaldehyde solution in Example 2 was prepared, 0.32 g of disodium edetate was added and stirred with a magnetic stirring bar until it was completely dissolved.

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Abstract

The present invention relates to a stable protein and / or amino acid detecting composition that can be used as a reagent for in situ detection, such as on surfaces. The invention also relates to a method for detecting protein and / or amino acid on surfaces using the composition and kits comprising the composition.

Description

technical field [0001] The present invention relates to stable protein and / or amino acid detection compositions which can be used as an in situ detection reagent, for example, on a surface for in situ detection. The present invention also relates to methods for detecting proteins and / or amino acids on surfaces using the above compositions and kits comprising such compositions. Background technique [0002] Detection of proteins, eg, of naturally intact proteins and / or their subgroups of amino acids and peptides, especially in trace amounts, is an important analytical and standard procedure in biotechnology in laboratory, clinical, and forensic settings. Typically, protein and / or amino acid detection is accomplished using off-the-shelf reagents, a combination of different techniques and instruments that have been validated. Existing methods tend to rely on reagents reacting with free amino acids and terminal amino acid residues of peptides, or reacting with terminal amino ac...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/52G01N33/68
CPCG01N33/52G01N33/6803G01N33/542G01N33/582G01N33/6839
Inventor 大卫·佩雷特索莱达·塞西莉亚·瑞兹·安拓丽南达·琪朔·巴布·南羽尼
Owner QUEEN MARY UNIV OF LONDON
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