Kit for detecting activity of human tissue kallikrein and application

A kallikrein and activity detection technology, applied in measurement devices, instruments, biomaterial analysis, etc., can solve problems such as unfavorable TK activity detection, complex protein composition, etc., and achieve high sensitivity, good repeatability, and strong specificity. Effect

Active Publication Date: 2014-09-17
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a human tissue kallikrein activity detection kit and its application. Combining the characteristics of the chemical reaction of BAEE to solve the problem that the complex protein composition in plasma is not conducive to the detection of TK activity

Method used

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  • Kit for detecting activity of human tissue kallikrein and application
  • Kit for detecting activity of human tissue kallikrein and application
  • Kit for detecting activity of human tissue kallikrein and application

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Embodiment Construction

[0041] In order to better explain the present invention, the main content of the present invention is further clarified below in conjunction with specific examples, but the content of the present invention is not limited to the following examples.

[0042] Preparation of Human Tissue Kallikrein Activity Detection Kit

[0043] 1. Reagents

[0044] 1) Coating buffer (pH=9.6 / 0.05 carbonate buffer):

[0045]

[0046] 2) PBST diluted washing solution (pH=7.4 / 0.15M)

[0047]

[0048] 3) BAEE working fluid

[0049]Dilute 1.7191 g of BAEE powder to 100 ml with Tris-HCl buffer solution at pH=8.0, which is 0.05M BAEE.

[0050] 4) Other reagents

[0051] Dilute hydrochloric acid is made from concentrated hydrochloric acid (concentration about 37%) and water in a volume ratio of 1:2, and its mass fraction is 13.73%.

[0052] The equivalent concentration of sodium hydroxide solution is 3.5N, the molar concentration of ferric chloride solution is 0.11M and the molar concentration...

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Abstract

A disclosed kit for detecting activity of human tissue kallikrein is characterized by comprising: 1) an enzyme label plate coated by specific anti-human tissue kallikrein clone antibody; 2) a standard sample of fusion protein TK-4T1 of human tissue kallikrein; and 3) a BAEE (N-benzoyl-L-arginine ethyl ester hydrochloride 0 working solution. According to the kit, a method combining immunization and chemistry is employed, by utilizing the characteristic that human tissue kallikrein (TK) in a human urine or blood plasma sample can be combined with the specific antibody, TK is separated from the sample and further catalyzes a chemical reaction of a substrate N-benzoyl-L-arginine ethyl ester hydrochloride (BAEE).

Description

technical field [0001] The invention relates to the field of human tissue kallikrein detection, in particular to a human tissue kallikrein activity detection kit and its application, which are used to detect the human tissue kallikrein activity. Background technique [0002] Human tissue kallikrein (TK) is a kind of serine protease, which can promote the release of vasoactive peptide by low-molecular-weight kininogen to exert a series of biological activities, named KLK1. In human and animal models of hypertension, cardiovascular disease and kidney disease, the reduction of TK content can be seen. We found that TK can lower blood pressure, improve insulin resistance, and improve renal function in transgenic animal models. The level of TK may have important predictive value in the occurrence and development of human cardiovascular disease and kidney disease. [0003] Our study found that the content of TK has a dose-response negative correlation with the occurrence of strok...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/543
CPCG01N33/543G01N33/573G01N2333/96433
Inventor 冉晓张琴
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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