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Novel signal amplification immunofluorescence kit

A signal amplification and immunofluorescence technology, applied in biological testing, material inspection products, measuring devices, etc., can solve the problems of insufficient amplification of weakly expressed protein targets, affecting the detection effect of markers, weak amplification results, etc., and achieve excellent detection effect, avoid autofluorescence interference, effect of low exposure value

Pending Publication Date: 2021-10-26
湖北百奥斯生物科技有限公司
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Problems solved by technology

However, the biotin-avidin amplification method will bring more background to the sliced ​​tissue, which will affect the detection effect of the label
[0006] The amplification effect of TSA technology alone is not ideal for weakly expressed protein targets, while the biotin-avidin amplification technology and the existing DAB amplification technology may bring more background or weak amplification results to the detected section tissue , especially when using fluorescent technology to detect weakly expressed proteins, it will also affect its detection effect. It is urgent to find an immunohistochemical detection kit and method with excellent signal amplification to solve the above-mentioned problems in the prior art

Method used

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Embodiment 1

[0066] A novel immunofluorescence signal amplification technique labeling method, comprising the following steps:

[0067] S0. Slicing: put the tissue slide in the slicing machine, set the temperature at 58°C for 2 hours;

[0068] S1. Dewaxing: Put the tissue slide into the clearing agent 10min successively, the clearing agent 10min, the clearing agent 10min, absolute ethanol 1 5min, 95% ethanol 5min, 75% ethanol 5min, rinse with tap water then, place in water ready to repair;

[0069] S2. Antigen restoration: Add EDTA of antigen restoration solution pH 8.0 to the pressure cooker, put the dewaxed and hydrated tissue slices into the buffer solution pH 5.0 PBS of the pressure cooker, wait for 1.5 minutes after the pressure cooker is sprayed, stop the fire; rinse with cold water Pour the pressure cooker, open the lid after the air pressure drops, and wait for the repair solution to cool down to room temperature naturally, take the slices out of the EDTA repair solution pH 8.0, s...

Embodiment 2

[0079]A novel immunofluorescence signal amplification technique labeling method, comprising the following steps:

[0080] S0. Slicing: put the tissue slide in the slicing machine, set the temperature at 59°C for 1.5 hours;

[0081] S1. Dewaxing: Put the tissue slide into the clearing agent 10min successively, the clearing agent 10min, the clearing agent 10min, absolute ethanol 1 5min, 95% ethanol 5min, 75% ethanol 5min, rinse with tap water then, place in water ready to repair;

[0082] S2. Antigen restoration: Add citric acid of antigen restoration solution pH 6.0 to the pressure cooker, put the dewaxed and hydrated tissue slices into the buffer solution pH 5.0 PBS of the pressure cooker, wait for 2 minutes after the pressure cooker is sprayed, stop the fire; rinse with cold water Pour the pressure cooker, and open the lid after the air pressure drops. After the repair solution cools down to room temperature naturally, take the slices out of the citric acid of the repair sol...

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Abstract

The invention provides an immunofluorescence kit for a novel signal amplification technology. The immunofluorescence kit specifically comprises the following components: a second antibody-HRP, a TSA-Alkyne reagent and an Azide-tracing marker. According to the kit, click chemistry and a TSA signal amplification method are combined, the advantages of the two methods are exerted, multi-round cyclic amplification can be performed according to experimental requirements, more backgrounds cannot be brought to slice tissues, the detection effect of a mark cannot be influenced, and when a fluorescence detection technology is used, the exposure value is low, autofluorescence interference can be avoided, and fluorescence signal detection can be more sensitive; and the kit has an excellent detection effect on a weak expression protein target spot.

Description

technical field [0001] The invention relates to the technical field of biomedical application, in particular to a novel signal amplification immunofluorescence kit. [0002] technical background [0003] Tyramide signal amplification (TSA, Tyramide signal amplification), also known as catalyzed signal amplification (CSA, catalyzed signal amplification), has been developed for nearly 20 years since its creation and is currently widely used. TSA technology is a kind of enzymatic detection method that uses HRP to carry out high-density in situ labeling of target antigens. It can not only be used for signal amplification of IF / IHC, but also for detection of Elisa and ISH. Compatible with high-performance dye Alexa Fluor, traditional fluorescent dyes, enzyme systems, and colorimetric detection systems. Its main principle is to use the peroxidase reaction of tyramide Tyramide (tyramide salt forms a covalent bond binding site under HRP catalysis H202), to produce a large number of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/531G01N33/53
CPCG01N33/582G01N33/531G01N33/5306
Inventor 陈志斌李晓芳肖鹏黄婷王培张瑞琦
Owner 湖北百奥斯生物科技有限公司
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