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Complement C1q determination kit and determination method thereof

A determination method and kit technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of inability to automate, high labor cost, and labor-intensive, and achieve the effect of improving detection sensitivity and detection sensitivity.

Pending Publication Date: 2020-09-22
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Disadvantages of the existing technology: At present, the detection of complement C1q has not been widely carried out, and most autoimmune diseases currently use the ELISA method, which requires a lot of manpower and cannot be automated, and the labor cost is high

Method used

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  • Complement C1q determination kit and determination method thereof

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Embodiment 1

[0038] A kit for determining complement C1q, the raw materials of which include R1 reagent and R2 reagent.

[0039] The present invention also provides an assay method of a complement C1q assay kit, comprising the following steps:

[0040] S1: Preparation of R2 reagent, the steps are as follows:

[0041] 1) Add 200ul of 133nm latex microspheres into a centrifuge tube, add 1700ul of 25mM MES solution, and mix for 25min at room temperature at 20°C;

[0042] 2) Add 1 mg of EDC and 3 mg of NHS, and mix for 12 minutes at 35°C;

[0043] 3) Centrifuge to remove the supernatant, add 1ml of 5mM CB solution, and ultrasonically resuspend;

[0044] 4) Dilute 0.8 mg of C1q antibody with 1 ml of 5 mM CB solution, quickly add it to the latex microspheres in step 2, and mix for 0.8 h at 35 ° C;

[0045] 5) In step 4, add 1ml of Glycine, 1ml of BSA and 3ml of Tris-HCL buffer to block overnight, and control the overnight blocking temperature at 2-8°C;

[0046] 6) Centrifuge the solution in ...

Embodiment 2

[0060] A complement C1q assay kit is characterized in that its raw materials are as follows: including R1 reagent and R2 reagent.

[0061] The present invention also provides an assay method of a complement C1q assay kit, comprising the following steps:

[0062] S1: Preparation of R2 reagent, the steps are as follows:

[0063] 1) Add 200ul of 133nm latex microspheres into a centrifuge tube, add 1700ul of 25mM MES solution, and mix for 30min at room temperature at 23°C;

[0064] 2) Add 1 mg of EDC and 3 mg of NHS, and mix for 15 minutes at 37°C;

[0065] 3) Centrifuge to remove the supernatant, add 1ml of 5mM CB solution, and ultrasonically resuspend;

[0066] 4) Dilute 0.8 mg of C1q antibody with 1 ml of 5 mM CB solution, quickly add it to the latex microspheres in step 2, and mix for 1 h at 37 ° C;

[0067] 5) In step 4, add 1ml of Glycine, 1ml of BSA and 3ml of Tris-HCL buffer to block overnight, and control the overnight blocking temperature at 2-8°C;

[0068] 6) Centri...

Embodiment 3

[0082] A complement C1q assay kit is characterized in that its raw materials are as follows: including R1 reagent and R2 reagent.

[0083] The present invention also provides an assay method of a complement C1q assay kit, comprising the following steps:

[0084] S1: Preparation of R2 reagent, the steps are as follows:

[0085] 1) Add 200ul of 133nm latex microspheres into a centrifuge tube, add 1700ul of 25mM MES solution, and mix for 35min at room temperature at 25°C;

[0086] 2) Add 1 mg of EDC and 3 mg of NHS, and mix for 18 minutes at 39°C;

[0087] 3) Centrifuge to remove the supernatant, add 1ml of 5mM CB solution, and ultrasonically resuspend;

[0088] 4) Dilute 0.8mg of C1q antibody with 1ml of 5mM CB solution, quickly add to the latex microspheres in step 2, and mix for 1.2h at 39°C;

[0089] 5) In step 4, add 1ml of Glycine, 1ml of BSA and 3ml of Tris-HCL buffer to block overnight, and control the blocking overnight temperature at 5°C;

[0090] 6) Centrifuge the ...

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Abstract

The invention discloses a complement C1q determination kit. The raw materials comprise a reagent R1 and a reagent R2, according to the invention, the complement C1q determination kit and the determination method thereof are strictly controlled; the proportion of each component of the complement C1q determination kit and preparation process parameters are strictly controlled; EDC and NHS are coupled with complement C1q antibodies and latex microspheres, the particle sizes of the latex microspheres are selected, polyethylene glycol or other polymerization promoters with different molecular weights are applied, and the C1q antibodies are polyclonal antibodies or a group of monoclonal antibodies or a combination of the monoclonal antibodies and the polyclonal antibodies. The latex microsphereswith larger particle sizes are selected, so that the detection sensitivity is also improved; by using high-molecular-weight polyethylene glycol or other polymerization promoters with higher polymerization promoting effects, the detection sensitivity is also improved.

Description

technical field [0001] The invention relates to the technical field of determination of complement C1q, in particular to a kit for determining complement C1q, and at the same time, the invention also relates to a method for determining the kit for determining complement C1q. Background technique [0002] C1q is an important component of complement C1, with a molecular weight of 390,000 and a symmetrical hexamer composed of six identical subunits. It is also the initiator of immune complexes through the classical (or traditional) pathway (CP) or activation of complement. When two or more C1qs combine with the Fc segment of IgM or IgG in the immune complex, the configuration of C1q changes, leading to the sequential activation of C1r and C1s, and the activation of the classical complement activation pathway. C1 in the circulation is a complex composed of 3 subunits, namely 1 C1q molecule, 2 C1r and 2 C1s molecules depending on the combination of Ca ions. [0003] Increased C1...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2333/4716
Inventor 程明陈平程心远胡敬生吕海沧杨柳张倩
Owner 程明
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