A kind of ring metal iridium complex and its preparation method and its application as protein staining agent

A technology of iridium complexes and ring metals, which is applied in the field of biological analysis, can solve problems such as complex rinsing steps, low dyeing sensitivity, and poor sensitivity of Coomassie Brilliant Blue, and achieve high dyeing sensitivity, simple preparation method, and good dyeing effect

Inactive Publication Date: 2019-01-29
SUZHOU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The dyes used in colorimetry include silver nitrate and Coomassie brilliant blue, etc., but silver nitrate and protein staining are combined through irreversible chemical bonds, which is not conducive to the subsequent analysis of proteins. Higher levels required; Coomassie Brilliant Blue less sensitive
The dyes used in the fluorescence method include two categories: small molecules and metal complexes. Metal complex luminescent dyes occupy an important position in the protein stain market due to their high stability and high luminous efficiency; however, the existing metal complexes Poor stability, low staining sensitivity, about a few hundred nanograms, and complicated washing steps

Method used

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  • A kind of ring metal iridium complex and its preparation method and its application as protein staining agent
  • A kind of ring metal iridium complex and its preparation method and its application as protein staining agent
  • A kind of ring metal iridium complex and its preparation method and its application as protein staining agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Synthesis of ionic luminescent ring metal iridium complexes

[0052]

[0053] First, after reacting iridium trichloride trihydrate crystalline compound with C^N bidentate ligand (compound 1) in ethylene glycol ether solvent for 24 hours, a transition metal chloride bridge compound (compound 2) was obtained, and then the chlorine bridge The compound was reacted with N^N bidentate ligand compound (compound 3), the solvent was spin-dried, and the final ionic luminescent ring metal iridium compound (compound 4) was obtained by column chromatography. Yield: 68%. Mass spectrum: experimental value [M-Cl] + 655.1610, theoretical value [M-Cl] + 655.1601.

[0054] By replacing the C^N bidentate ligand and N^N bidentate ligand compounds, ion-type luminescent ring metal iridium complexes with multiple structures can be obtained.

[0055] It is also possible to combine the chloride ion iridium complex obtained above with NaBr, KI or KPF 6 The reaction yields cyclom...

Embodiment 2

[0056] Example 2 Preparation of electrically neutral and luminescent ring metal iridium complexes

[0057]

[0058] Iridium acetylacetonate Ir(acac) 3 Heated and reacted with C^N bidentate ligand compound 1 for 24 hours, and separated by column chromatography to obtain electrically neutral luminescent ring metal iridium complex 2. The yield was 60%. Mass Spectrum: Experimental [M+H] + 655.1510, theoretical value [M+H] + 655.1599.

[0059] By replacing the C^N bidentate ligand compound, multiple structures of electrically neutral luminescent ring metal iridium complexes can be obtained.

Embodiment 3

[0061] The structural formula of the protein stain used is:

[0062]

[0063] The fluorescence quantum efficiency of this compound is 37%.

[0064] The ring metal iridium complex of the present invention is used for protein staining and gel test, the process is as follows:

[0065] 1. After the protein sample was subjected to SDS-PAGE, it was fixed in 30% ethanol and 10% acetic acid;

[0066] 2. Rinse the glue in 20% ethanol for 3 times, 30 minutes each time;

[0067] 3. Dilute the protein dye luminescent iridium complex from the mother liquor (with DMSO as the solvent) to 1 micromol with pure water, and then place the gel in it for staining for 6 hours;

[0068] 4. Take out the gel after the third step and place it in the gel imaging system for imaging. Choose 312nm as excitation wavelength and 515nm as emission wavelength.

[0069] Stain commercially available protein markers (molecular weight: 116KDa, 66.2 KDa, 45 KDa, 35 KDa, 25 KDa, 18.4 KDa, 14.4 KDa) separated by...

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Abstract

The invention discloses a ring metal iridium coordination compound, a preparation method thereof, and an application of the ring metal iridium coordination compound as a protein dyeing agent. The ring metal iridium coordination compound comprises a C^N bidentate ligand and / or an N^N bidentate ligand structure, belongs to a cationic type light-emitting metal coordination compound or an electrically neutral light-emitting metal coordination compound, and has a high quantum efficiency. When being used as the protein dyeing agent, the ring metal iridium coordination compound is high in dyeing sensitivity, ng levels can be reached, and a rinsing step is not needed during dyeing, so that more time is saved; the problems that rinsing is needed, and the sensitivity is low in the prior art are solved; and besides, the ring metal iridium coordination compound disclosed by the invention is simple in preparation method and high in stability.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a cyclometal iridium complex, a preparation method thereof and an application as a protein stain. Background technique [0002] As an important technique, electrophoresis is widely used in biologically related fields such as protein analysis and drug development. By means of electrophoresis, proteins can be separated into different bands on sodium dodecylsulfonate-polyacrylamide gel plates (SDS-PAGE) according to different molecular weights. The visualization of these protein bands is the premise and basis for subsequent protein analysis. [0003] Protein visualization methods currently include colorimetric and fluorescent methods. The dyes used in colorimetry include silver nitrate and Coomassie brilliant blue, etc., but silver nitrate and protein staining are combined through irreversible chemical bonds, which is not conducive to the subsequent analysi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07F15/00G01N1/30C09K11/06
CPCC07F15/0033C09K11/06C09K2211/185G01N1/30
Inventor 周宇扬贾俊丽
Owner SUZHOU UNIV OF SCI & TECH
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