Quantifying device for human neutrophil lipophorin homodimers

A technology of homodimer and neutrophils, which is applied in the field of immunology detection, can solve the problems of complex operation, difficulty in accurate quantification, and long test cycle, and achieve the effect of simple preparation method, low detection limit, and quick use

Inactive Publication Date: 2016-09-28
JILIN UNIV
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  • Application Information

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Problems solved by technology

The above-mentioned HNL quantitative methods have their own advantages, disadvantages and corresponding uses. However, except for Western-blotting, which can distinguish different molecular forms of HNL, none of the above-mentioned HNL quantitative methods can specifically quantify homodimer HNL, which affects the use of this protein in some diseases. suc...

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  • Quantifying device for human neutrophil lipophorin homodimers
  • Quantifying device for human neutrophil lipophorin homodimers
  • Quantifying device for human neutrophil lipophorin homodimers

Examples

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Embodiment 1

[0058] Embodiment 1: The composition of a kind of quantitative device of human neutrophil apolipoprotein homodimer

[0059] The device of this embodiment contains the following reagents and materials (see Table 3).

[0060] Table 3: The composition of embodiment 1 device

[0061] serial number

Embodiment 2

[0062] Embodiment 2: The composition of a kind of quantitative device of human neutrophil apolipoprotein homodimer

[0063] The device of this embodiment contains the following reagents and materials (see Table 4).

[0064] Table 4: The composition of embodiment 2 device

[0065] serial number

Embodiment 3

[0066] Example 3 Preparation of a 96-well ELISA plate coated with an anti-HNL antibody for a quantitative device for human neutrophil apolipoprotein homodimer

[0067] The preparation steps of the 96-well microtiter plate coated with the anti-HNL antibody are illustrated by taking the 96-well microtiter plate coated with the anti-HNL antibody included in Example 1 as an example. Specific steps are as follows:

[0068] (1) Anti-HNL antibody coated 96-well ELISA plate

[0069] Anti-HNL monoclonal antibody (ab23477 of Abcam company) was used 50mmol / LNa 2 CO 3 -NaHCO 3 , pH 9.6 sodium carbonate buffer diluted to 1 μg / mL to make antibody coating solution; to 96-well enzyme plate (Nunc flat-bottom 96well plate) Add 100 μL of antibody coating solution to each well; overnight at 4°C.

[0070] (2) Sealed microtiter plate

[0071] Discard the antibody coating solution; add 300 μL of blocking solution to each well of the 96-well ELISA plate (the blocking solution is 50 mmol / L Na ...

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Abstract

The invention relates to a quantitative device for human neutrophil apolipoprotein (HNL) homodimer, which belongs to the technical field of immunological detection. Consists of 5-fold sample dilution, 25-fold washing solution, HNL homodimer standard stock solution, 96-well enzyme-labeled plate coated with anti-HNL antibody, enzyme-labeled anti-HNL antibody, U-bottom 96-well plate, sealing film , enzyme substrate solution and stop solution. The anti-HNL antibody coated on the enzyme plate of the present invention and the enzyme-labeled anti-HNL antibody are the same antibody, thereby simply realizing the quantification of the HNL homodimer concentration. In addition, only two steps of incubation and one step of washing are required during the use of the device, thereby significantly shortening the detection time. Therefore, the invention has the advantages of simple preparation, convenient use, short test time and the like. The invention can be used for the detection and monitoring of diseases (such as acute bacterial infection) marked by HNL homodimer, and thus has great laboratory research and clinical application value.

Description

technical field [0001] The invention belongs to the technical field of immunological detection, and in particular relates to a rapid quantitative device for human neutrophil apolipoprotein (HNL) homodimer. Background technique [0002] Human neutrophil lipocalin (human neutrophil lipocalin, HNL) is also called neutrophil gelatinase associated lipocalin (neutrophil gelatinase associated lipocalin, NGAL), also known as lipocalin-2 (Lipocalin-2 ), which is a lipocalin (J. Biol. Chem. 1993, 268: 10425-10432; Scand. J. Clin. Lab. Invest. 1994, 54: 365-376). HNL exists in various molecular forms such as monomer, homodimer and heterodimer (formation of monomer and MMP9 molecule), and different molecular forms of HNL are related to different pathological processes. For example, monomeric HNL may have a relatively high correlation with acute kidney injury (Clin.Chim.Acta.2009,403:121-125; Lancet 2005,365:1231-1238; Biomed.Res.Int.2015,2009, 186:48-51; Am.J.Nephrol.2004,24:307-315; ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893
Inventor 蔡林君姜春来孔维
Owner JILIN UNIV
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