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Coating buffer solution, kit and application

A technology for coating buffers and buffers, applied in the field of clinical diagnosis, can solve problems such as low detection sensitivity and low P/N, and achieve the effects of solving low sensitivity, improving P/N, and improving sensitivity and specificity

Active Publication Date: 2020-06-05
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, the object of the present invention is to solve the problems of low P / N and low detection sensitivity in the enzyme-linked immunosorbent assay method existing in the prior art, and provide a method that can improve the positive sample signal and negative sample signal of the detection test. Coating Buffer for Signal Ratio (P / N) ELISA

Method used

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  • Coating buffer solution, kit and application
  • Coating buffer solution, kit and application
  • Coating buffer solution, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, preparation of different coating buffers and coating liquid

[0025] 1. Preparation of different coating buffers

[0026] 1. Prepare 100mL Tris coating buffer: weigh 0.242g of Tris in a beaker, measure 55mL of purified water with a graduated cylinder, pour it into the beaker, put it into the stirring rotor, add 0.1mL of Proclin300, put it on a magnetic stirrer and stir until completely dissolve. Measure the pH of the above solution with a calibrated pH meter, adjust the pH to 8.50±0.20 with 2M HCl or 2M NaOH, and set the volume to 100 mL.

[0027] 2. Prepare 100mL LCB coating buffer: weigh Na 2 CO 3 0.159g, NaHCO 3 0.293g, add 0.1mL of ProClin 300, measure 55mL of purified water with a graduated cylinder, pour it into a beaker, put it into a stirring rotor, and stir on a magnetic stirrer until completely dissolved. Measure the pH of the above solution with a calibrated pH meter, adjust the pH to 9.60±0.20 with 2M HCl or 2M NaOH, and set the volume t...

Embodiment 2

[0044] Embodiment 2, the P / N value comparison that adopts different coating liquids to detect positive reference substance and negative reference substance

[0045] 1. Coating of 96-well plate (Nunc, catalog number: 446469)

[0046] Dilute the THSD7A-P1 protein with the prepared 14 coating buffers to a concentration of 0.4 μg / mL, coat 100 μL per well, put the coated plate in the refrigerator (2-8°C environment), and coat for 16-20 hours.

[0047] 2. The coating plate is closed and stable

[0048] 2.1 Dosing

[0049] 10× Concentrated Wash Solution 100mL: Weigh 2.422g of NaCl and 8.766g of Tris and dissolve them in purified water, add 110μL of Proclin300 and 50μL of Tween 20, mix well, adjust the pH to 7.2±0.2, and dilute to 100mL with purified water.

[0050] PBS1000mL: Weigh Na 2 HPO 4 12H 2 O 2.9006g, NaH 2 PO 4 2H 2 O 0.2964g, NaCl 8.766g, adjust the pH to 7.2±0.2 and dilute to 1000mL with purified water.

[0051] 2.2 Washing solution to wash the plate

[0052] Dil...

Embodiment 3

[0081] Example 3: The THSD7A antibody detection kit was prepared with PBS coating buffer and PBS-TGP1 coating buffer respectively, and the detection of negative and positive clinical samples was carried out.

[0082] 1. Coating of 96-well plate (Nunc, catalog number: 446469)

[0083] The preparation of PBS and PBS-TGP1 coating buffer is the same as that in Example 1. Dilute THSD7A-P1 protein with PBS coating buffer and PBS-TGP1 coating buffer respectively to a concentration of 0.4 μg / mL, coat 100 μL per well, put the coated plate in the refrigerator (at 2-8°C), and coat By 16-20h.

[0084] 2. The coating plate is closed and stable

[0085] 2.1 Dosing

[0086] 10× Concentrated Wash Solution 100mL: Weigh 2.422g of NaCl and 8.766g of Tris and dissolve them in purified water, add 110μL of Proclin300 and 50μL of Tween 20, mix well, adjust the pH to 7.2±0.2, and dilute to 100mL with purified water.

[0087] PBS1000mL: Weigh 2.9006g of Na2HPO4·12H2O, 0.2964g of NaH2PO4·2H2O, 8.76...

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Abstract

The invention belongs to the technical field of clinical diagnosis, and discloses a coating buffer solution, a kit and application. The coating buffer solution provided by the invention comprises a buffer salt solution and 5v / v%-15v / v% glycerol. By adopting the coating buffer solution to dilute a sample to be detected and then coating an elisa plate for ELISA detection, P / N can be obviously improved, the detection sensitivity and specificity are further improved, the problem of low sensitivity is solved, and the positive coincidence rate and the negative coincidence rate of a detection test can both reach 100%. The coating buffer solution or the kit comprising the coating buffer solution can be used for preparing THSD7A-P1 protein diagnosis products.

Description

technical field [0001] The invention belongs to the technical field of clinical diagnosis, and in particular relates to a coating buffer, a kit and its application, in particular to a THSD7A antibody detection kit. Background technique [0002] Idiopathic Membranous Nephropathy (Idiopathic Membranous Nephropathy, IMN) is a group of diseases characterized by deposition of immune complexes under the glomerular basement membrane epithelial cells and diffuse thickening of the basement membrane. One of the common types of kidney disease, the age of onset is 50-60 years old. In recent years, the incidence of this disease has gradually increased and younger. Typical clinical features of IMN are massive proteinuria (≥3.5 g / 24-hour urine), hypoalbuminemia (<30 g / dL), hyperlipidemia, and edema. Studies have shown that nearly 40% of all IMN patients will eventually enter chronic renal failure. For IMN patients, although there are some clinical indicators that can guide clinicians ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53
CPCG01N33/6893G01N33/5306G01N2800/347G01N2333/47
Inventor 王洪涛张大准詹婷婷梁香禄
Owner SHENZHEN BLOT BIOTECH