A kind of dual-mode ELISA enzyme immunochromogenic reagent and its preparation and application
A dual-mode, chromogenic agent technology, applied in the field of immunology and immunoassay, can solve the problems of increasing experimental operation steps and sacrificing the simplicity of traditional ELISA, and achieves high detection sensitivity, excellent optical stability, and fast enzyme response speed and the effect of sensitivity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0047] In this embodiment, the dual-mode ELISA enzyme-immune chromogenic reagent includes the aggregation-induced luminescent material TPE-(NH 2 ) 4 . The chromogen also includes hydrogen peroxide and PBS buffer solution.
[0048] The preparation of the dual-mode ELISA enzyme immunochromogenic reagent and its color development method are as follows:
[0049] (1) TPE-(NH 2 ) 4 Dissolve in 1,4-dioxane, the final concentration is 10mM;
[0050] (2) Dissolve HRP in 1×PBS buffer, the final concentration is 1mg / mL;
[0051] (3) Configure H 2 o 2 Solution, the concentration is 0.1M;
[0052] (4) Take 30μL TPE-(NH 2 ) 4 solution, add 70 μL 1×PBS buffer solution, add 10 μL hydrogen peroxide solution, and mix well to obtain a dual-mode ELISA enzyme immunochromogenic reagent solution; then add 10 μL HRP solution and react for 30 minutes; repeat the same operation above 10 times, measure each solution Absorption spectrum, take the intensity of the absorption value at 600nm to m...
Embodiment 2
[0056] (1) Take a 96-well plate, add 70 μL 1×PBS, 30 μL TPE-(NH 2 ) 4 solution, from left to right, adding TPE-(NH 2 ) 4 The solution concentrations were 10mM, 1mM, 100μM, 10μM, 1μM, 100nM, 10nM, 1nM; then 10μL HRP (1mg / mL) and 10μL H 2 o 2 (0.1M);
[0057] In another line, add 70 μL 1×PBS, and then add 30 μL TPE-(NH 2 ) 4 solution (10mM), followed by adding 10μL H 2 o 2 , and finally add 10 μL of different concentrations of HRP, from left to right, the HRP concentrations are 2mg / mL, 1mg / mL, 100μg / mL, 10μg / mL, 1μg / mL, 100ng / mL, 10ng / mL, 1ng / mL;
[0058] In another row, add 70 μL 1×PBS, 30 μL TPE-(NH 2 ) 4 solution (10mM), followed by adding 10μL of different concentrations of H 2 o 2 , from left to right, H 2 o 2 Concentrations were 100mM, 10mM, 1mM, 100μM, 10μM, 1μM, 100nM, 10nM, and finally 10μL of HRP (1mg / mL) was added to each well.
[0059] (2) Take a 96-well plate, add 70 μL 1×PBS to the four rows of wells, add 30 μL OPD (10 mM) to the first row of wells, ...
Embodiment 3
[0065] (1) Prepare glucose oxidase solution (1×PBS, 1mg / mL), glucose solution (0.1M); take 70μL 1×PBS, add 30μL TPE-(NH 2 ) 4 solution, followed by adding 10 μL of glucose solution, glucose oxidase solution (10 μL, 2 mg / mL), and finally adding 10 μL of HRP enzyme. After reacting for 30 minutes, stop the reaction with 100 μL of deionized water.
[0066] (2) Take 10 μL of gold nanoparticles (15nm, 10nM,) solution, add 10 glucose solution (0.1M), react for half an hour, centrifuge to remove gold nanoparticles, keep the supernatant; take another test tube, add 70 μL to it successively 1×PBS, 30 μL TPE-(NH 2 ) 4 10 μL of HRP enzyme solution (1 mg / mL) was finally added to the gold nanocatalytic supernatant obtained above, and after 2 hours of reaction, the reaction was terminated with 100 μL of deionized water.
[0067] (3) DNA and Hemin form a G-quadruplex / Hemin complex structure in solution, the concentration of DNA is 100nM, the concentration of Hemin is 200nM, and the buffer ...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Sensitivity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More