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Chemiluminiscence substrate detection kit with high sensitivity and stable signals

A detection kit and high-sensitivity technology, applied in chemiluminescence/bioluminescence, biological testing, and analysis through chemical reactions of materials, can solve the problems of fast attenuation, strong signal strength, and high price, and achieve Long duration, high sensitivity, and the effect of maintaining signal stability

Pending Publication Date: 2021-01-08
上海雅酶生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the reaction catalyzed by 4-aminopyridine is difficult to control, even a very small amount of compound will generate a large amount of light signal, the signal intensity is strong, but the decay speed is also fast
[0005] Imported kits have products with strong signal and long duration of signal, but the price is very expensive

Method used

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  • Chemiluminiscence substrate detection kit with high sensitivity and stable signals
  • Chemiluminiscence substrate detection kit with high sensitivity and stable signals
  • Chemiluminiscence substrate detection kit with high sensitivity and stable signals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1: the influence of purine enhancer on luminol chemiluminescence;

[0067] Step 1: Configure reagent A and reagent B;

[0068] Reagent A: sodium perborate tetrahydrate 10mM

[0069] Citrate buffer pH 5.0 0.01M

[0070] Reagent B: Luminol sodium salt 5mM

[0071] Sodium 3-(phenothiazin-10-yl)propane 1-sulfonate 5mM

[0072] Co-enhancers:

[0073] (1) No co-enhancer-

[0074] (2) 4-N’N’. Dimethylaminopyridine (DMAP) 10mM

[0075] (3) Adenine: 10mM

[0076] (4) Purine: 10mM

[0077] N-cyclohexyl-2-aminoethanesulfonic acid pH9.0 100mM

[0078] Step 2: Store the prepared reagent A and reagent B in a refrigerator at 4°C. Mix reagent A and reagent B in equal proportions during use;

[0079] Step 3: Disposable polystyrene black 96-well microplate. Add 200ul of the above example working solution to each well of the microplate, then add 2ul of 100ng / mL horseradish peroxidase solution (HRP type VIa) to each well, and mix well. Using the Synergy-H1 full-featu...

Embodiment 2

[0081] Embodiment 2: Western Blot detection;

[0082] The kit of the invention is applied to the immunoassay analysis of HRP markers.

[0083] Reagent A: sodium perborate tetrahydrate 8mM

[0084] Citrate buffer pH 5.0 10mM

[0085] Reagent B Luminol Sodium Salt 2mM

[0086] Sodium 3-(phenothiazin-10-yl)propane 1-sulfonate 5mM

[0087] Adenine 8mM

[0088] N-cyclohexyl-2-aminoethanesulfonic acid pH9.1 100mM

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Abstract

The invention discloses a chemiluminiscence substrate detection kit with high sensitivity and stable signals, and relates to the technical field of biomedical kits. The kit comprises a reagent A and areagent B, wherein the reagent A is an oxide prepared from any one of hydrogen peroxide, sodium perborate or urea peroxide, a buffer substance and a stabilizer; the reagent B is a chemiluminescent substrate and comprises a luminescent agent adopting any one of luminol or luminol sodium salt or luminol isomer, a reinforcing agent adopting any one of n-alkyl phenothiazine or sodium salt thereof, and a second enhancer. The chemiluminiscence intensity can be improved, and the signal stability can be maintained; the detection kit has the advantages of being high in sensitivity, long in chemical signal lasting time, suitable for fluorescence scanning of a fluorescence CCD camera and capable of sensing an X-ray film and has chemical signal stabilization and good experiment repeatability.

Description

technical field [0001] The invention belongs to the technical field of biomedical kits, in particular to a chemiluminescent substrate detection kit with high sensitivity and stable signals. Background technique [0002] Chemiluminescence immunoassay (CLIA) is a detection method that combines chemiluminescence technology with immunochemical reactions, and can be used for the detection and analysis of antigens, antibodies, nucleic acids, etc., in immunoblotting such as Western Blot, DNA It is widely used in Southern Blot and Northern Blot. [0003] Horseradish peroxidase (HRP) is often used to label antigens or antibodies. After the immune reaction, luminol is used as a luminescent substrate, which emits light through redox reactions under the action of enzyme-labeled immunoreactants and luminescent reagents, and then according to the luminescence intensity Identify antigens or antibodies. However, as a luminescent substrate, luminol has poor stability, short signal duration...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/535G01N21/76
CPCG01N33/5306G01N33/535G01N21/76
Inventor 吴军黄人卉赵剑
Owner 上海雅酶生物医药科技有限公司
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