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CK-MB detection kit and use method thereof

A detection kit, tween-20 technology, applied in the field of immunochemical detection, can solve problems such as unrealizable, achieve the effect of improving detection speed, improving stability, and satisfying rapid diagnosis

Active Publication Date: 2018-04-24
NANTONG EGENS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defect that the CK-MB detection in the prior art cannot realize the detection of whole blood samples

Method used

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  • CK-MB detection kit and use method thereof

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Embodiment 1

[0057] This embodiment provides a method for preparing an alkaline phosphatase-labeled CK-MB antibody, comprising the following steps:

[0058] 1. Weigh an appropriate amount of Traut's reagent, and prepare a 1.376 mg / mL solution with pH 8.5, 100 mM triethanolamine buffer.

[0059] 2. Store the CK-MB labeled antibody (that is, the CK-MB antibody) in pH 8.5, 100mM triethanolamine buffer, and the antibody concentration is 2mg / mL; add the Traut' prepared in step 1 to the above antibody solution s solution, the molar ratio of Traut's reagent and CK-MB antibody is 30:1, mix immediately, and stand at 25°C for 15 minutes.

[0060] 3. Add 1M glycine solution to the antibody solution obtained after the activation reaction in step 2, and the molar ratio of glycine to Traut's reagent is 20:1; mix immediately, and stand at 25°C for 10 minutes for reaction.

[0061] 4. The CK-MB labeled antibody solution obtained after the reaction in step 3 was immediately desalted, replaced with a pH 8....

Embodiment 2

[0072] This embodiment provides a method for preparing an alkaline phosphatase-labeled CK-MB antibody, comprising the following steps:

[0073]1. Weigh an appropriate amount of Traut's reagent, and prepare a 1.376 mg / mL solution with pH 8.5, 100 mM triethanolamine buffer.

[0074] 2. Store the CK-MB labeled antibody (that is, the CK-MB antibody) in a pH 8.5, 100 mM triethanolamine buffer solution with an antibody concentration of 5 mg / mL; add the Traut' prepared in step 1 to the above antibody solution s solution, the molar ratio of Traut's reagent to CK-MB antibody is 15:1, mix immediately, and stand at 25°C for 15 minutes.

[0075] 3. Add 1M glycine solution to the antibody solution obtained after the activation reaction in step 2. The molar ratio of glycine to Traut's reagent added is 10:1; mix immediately, and stand at 25°C for 10 minutes for reaction.

[0076] 4. The CK-MB labeled antibody solution obtained after the reaction in step 3 was immediately desalted, replaced ...

Embodiment 3

[0087] This embodiment provides a method for preparing CK-MB antibody-labeled magnetic beads, comprising the following steps:

[0088] 1. Store the CK-MB coating antibody (that is, CK-MB antibody) in MES buffer solution with pH 5.0 and 100mM, and the antibody concentration is 1mg / mL;

[0089] 2. Take the carboxyl magnetic bead solution 100 times the weight of the antibody, the concentration of the carboxyl magnetic bead solution is 100 mg / ml, the particle size of the carboxyl magnetic bead is 3 μm, and the supernatant is discarded by magnetic separation;

[0090] 3. Washing: re-dissolve the magnetic beads obtained in the above steps with pH 5.0, 100 mM MES buffer, and discard the supernatant by magnetic separation.

[0091] 4. Repeat step 3 once.

[0092] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.

[0093] 6. Weigh an appropriate amount of NHS (N-hydroxysuccinimide) reagent and dissolve it with pH 5.0, 100mM MES buffer...

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Abstract

The invention discloses a CK-MB detection kit which comprises calibrator, cleaning fluid, a substrate solution, a pretreatment solution, an enzyme conjugate working solution and magnetic bead conjugate working solution, wherein the pretreatment solution contains imidazole, the enzyme conjugate working solution contains an enzyme marked CK-MB antibody, and the magnetic bead conjugate working solution contains CK-MB antibody marked magnetic beads. By means of the CK-MB detection kit, accurate measurement of CK-MB in a whole blood sample can be achieved, detection steps are simplified, and a detection efficiency is improved. The lowest detection limit of the kit is 1ng / ml, a linearity range is 1 to 300ng / ml, detection sensitivity is high, the linearity range is wide, and detection results areaccurate. The invention further discloses a use method of the CK-MB detection kit. Use steps of the use method are simple, CK-MB detection time is effectively shortened, and quick and flexible detection of the CK-MB is benefit to achieve.

Description

technical field [0001] The invention belongs to the technical field of immunochemical detection, and in particular relates to a CK-MB detection kit and a use method thereof. Background technique [0002] Cardiovascular disease is a major fatal disease that endangers the health and life of the people in our country. According to the statistics of the Ministry of Health, in my country's cities, 235 people die of cardiovascular and cerebrovascular diseases for every 100,000 people, accounting for 100% of all diseases. The number of deaths caused by the disease is the first, and it is increasing at a rate of 2% every year. Acute Myocardial Infarction (AMI) is the main cause of death of cardiovascular patients. AMI is a clinically sudden disease. Its onset is often accompanied by a series of symptoms such as chest pain, nausea, fever, arrhythmia, and elevated serum markers. Early diagnosis and interventional treatment will help save dying myocardium and improve prognosis. It is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/535G01N33/543
CPCG01N33/535G01N33/54326G01N33/573
Inventor 王保君汤双双欧卫军沙海邵志燕
Owner NANTONG EGENS BIOTECH CO LTD
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