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Protein chip detection kit

A detection kit and protein chip technology, applied in biological testing, material inspection products, etc., can solve the problems of low signal detection rate, low detection signal, and undisclosed components, and achieve high signal detection rate and signal detection. Strong, low-background effect

Inactive Publication Date: 2018-01-09
WAYEN BIOTECHNOLGIES SHANGHAI INC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most protein chip companies at home and abroad have special detection kits suitable for the protein chips produced by the company, but the kits are expensive, the ingredients are not disclosed, and they are not suitable for protein chips prepared by other types of surface chemically modified substrates. The detection, specifically manifested in the shortcomings of low detection signal
However, traditional chip hybridization reagents, such as hybridization buffers containing different proportions of Tween, skimmed milk powder or BSA, have disadvantages such as high chip background and low signal detection rate.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0013] Embodiment 1 protein chip detection kit preparation process

[0014] A. Hybridization solution: 0.75g sodium chloride, 30g glycine, 0.23g potassium dihydrogen phosphate, 0.47g disodium hydrogen phosphate, 10.00g sucrose, 10ml 2% EDTA, 500ul Tween-20, add Set the volume to 1000ml with ionized water, adjust the pH to 7.0, take an appropriate amount and add 3% skimmed milk powder to mix well before use.

[0015] B. Blocking solution: made by adding 0.05% Tween-20 to 0.01M phosphate buffer solution, the specific method is: weigh 8.00g sodium chloride, 0.2g potassium chloride, 1.44g disodium hydrogen phosphate, 0.24g phosphoric acid Dipotassium hydrogen, 500ul Tween-20, dilute to 1000ml, add 3% skimmed milk powder and mix well before use.

[0016] C. Cleaning solution: made by adding 0.05% Tween-20 to 0.01M phosphate buffer solution, the specific method is: weigh 80.00g sodium chloride, 2.00g potassium chloride, 14.40g disodium hydrogen phosphate, 2.40g phosphoric acid Dip...

Embodiment 2

[0018] Example 2 Using the kit of the present invention to detect the reaction of cell lysate and protein chip

[0019] (1) Protein chip preparation: 21 kinds of antibodies (purchased from immunoway) and 2 kinds of ginseng (biotin-anti-Mouse) were spotted on the chip using the sciFLEXARRAYER S1 chip spotter, and the chip substrate was modified with 3 different surface chemicals , were silylated siliconized film base and hydrophilic polymer modified film base, which were purchased from SCHOTT, Germany, and aldehyde-modified film base, which were purchased from Beijing Boao Biotechnology. 4 replicates were spotted for each antibody, and 2 quadruple replicates (located at the upper left and lower right of the reaction area of ​​the chip respectively) were spotted for each positive ginseng to form an independent reaction area. The antibody layout is shown in Table 1. The protein names corresponding to the antibodies are shown in Table 2.

[0020] Table 1. Antibody arrangement fo...

Embodiment 3

[0037] Example 3 Using the kit of the present invention to detect the reaction between the tissue lysate and the protein chip

[0038] Referring to the operation process of Example 2, 25ug of biotin-labeled mouse liver tissue lysate was used to hybridize with three types of chip-based chips, and the kit and FM kit of the present invention were used for detection. The test results are shown in Table 4.

[0039] Table 4. Test results of tissue lysates.

[0040]

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Abstract

The invention provides a protein chip detection kit. The protein chip detection kit comprises hybridization solution A, confining liquid B, washing liquid C and skimmed milk powder D. The protein chipdetection kit provided by the invention has the characteristics of being easy in operation, high in signal detection strength, high in signal detection rate and low in background and is applicable toprotein chips prepared by utilizing multiple surface chemically modified film bases, and an effective low-cost detection kit is provided for detection of the protein chips.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a protein chip detection kit. Background technique [0002] Protein chip is a high-throughput detection technology, which is a biological detection technology developed in recent years. Using this technology, the quantitative changes of multiple proteins can be detected simultaneously by placing a large number of antibodies densely on a solid-phase carrier. The technique is efficient and requires a small sample size. The more important step in the protein chip detection technology process is the hybridization reaction between the sample and the chip, which has a great impact on the detection results of the protein chip, mainly reflected in the chip background, signal detection rate, signal intensity, and signal uniformity. [0003] At present, most protein chip companies at home and abroad have special detection kits suitable for the protein chips produced by the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/68
Inventor 张庆华宋凯李文娜
Owner WAYEN BIOTECHNOLGIES SHANGHAI INC
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