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Method for determining microprotein in near infrared wavelength zone based on dye binding principle

A technology in the near-infrared and wavelength range, which is applied in the field of protein determination, can solve the problem of no substantial improvement in sensitivity, and achieve the effects of high sensitivity, large molar absorption coefficient, and significant enhancement of absorbance

Inactive Publication Date: 2004-12-15
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Numerous methods have also been published for protein quantitation using dye-conjugated methods, but with no substantial improvement in sensitivity over Coomassie Brilliant Blue

Method used

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  • Method for determining microprotein in near infrared wavelength zone based on dye binding principle
  • Method for determining microprotein in near infrared wavelength zone based on dye binding principle
  • Method for determining microprotein in near infrared wavelength zone based on dye binding principle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0044] An example of experimental operation under optimized conditions is given below.

[0045] In a series of 10mL volumetric flasks, 0.0, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2mL of human serum albumin (HSA) aqueous solution with a concentration of 20ug / mL were added in sequence, and then 1.0mL pH2.6 was added in sequence of glycine-hydrochloric acid buffer solution, dilute to full volume with water and shake well. Then add 0.3mL of 1.0×10 bottle by bottle -4 mol / L cyanine solution, shake well. After standing for 40 min, scan the absorption spectrum with the buffer solution as a reference or measure the absorbance at 904 nm. The parameters of each component are shown in the table below

[0046]

[0047] The response of the present invention to proteins under other conditions is given below.

[0048] 1) Response of different pH systems to proteins

[0049] Final BSA concentration (ng / mL)

Absorbance

pH 1.0 a

pH 4.0 b

0

0....

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Abstract

A method for detecting microprotein in near-infrared region based on dye binding principle includes adding the aqueous solution of protein in bottle with incremental concentrations, adding acidic buffer solution, diluting with water, adding cyanine solution, standing and scanning absorption spectrum or measuring the increment of light absorption degree. Its advantages are high sensitivity and less interference.

Description

(1) Technical field [0001] The invention relates to a method for measuring protein, in particular to a method for measuring trace protein in the near-infrared wavelength region based on the principle of dye binding. (2) Background technology [0002] Cyanine dyes belong to a class of compounds, which were first used as photosensitizers in the photographic chemical industry, or as energy conversion and storage materials, such as pigments for solar cells or laser dyes. In recent years, it has been found that these compounds have long-wave fluorescence (600-1000 nm) emission, so they are gradually applied in fluorescence analysis, combining laser-induced fluorescence (LIF) technology and capillary electrophoresis (Caterpillar electroporesis, CE) technology or chromatographic separation technology, Such compounds have been used for nucleic acid sequencing (Shealy, D.B. et al. Anal. Chem., 1995, 67, 247; Shealy, D.B. et al. Appl. Spectrosc., 1995, 49, 1815) and proteins (Sauda, ​...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/3577G01N21/359G01N33/52
Inventor 郑洪李东辉毛玉霞
Owner XIAMEN UNIV
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