Methods and reagent kits for determining activity of thioredoxin reductase and uses thereof
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A technology of thioredoxin and reductase, which is applied in the field of enzyme activity detection, can solve the problems of poor specificity and low sensitivity, and achieve the effect of accurate, accurate and sensitive anti-disease measurement results
Active Publication Date: 2014-05-14
KEAISE MEDICINE WUHAN +1
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Problems solved by technology
[0008] Therefore, it is urgent to solve the problems of low sensitivity and poor specificity in the detection of TrxR activity in samples.
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Embodiment 1
[0067] 1. Determination conditions of microplate reader
[0068] In the mode controlled by Ascent Software Version2.6,
[0069] Oscillation: time 5s, speed 1020rpm;
[0070] Measurement mode: continuous;
[0071] Measurement Type: Dynamic;
[0072] Time interval: 30s;
[0073] Number of measurements: 15;
[0074] Wavelength: 405nm.
[0075] When analyzing an actual blood sample, a standard curve is established for each analysis run to calculate the concentration of the analyte in the analysis run.
[0076] 2. Measurement process
[0077] 2.1 Preparation of working solution: take 1M K 2 HPO 4 615ml, 1M KH 2 PO 4 Mix 385ml, adjust the pH to 7.0, add 2924mg EDTA and 200mg BSA, dissolve, and store at 4°C.
[0078] 2.2 Preparation of inhibitor working solution (25 μl): Dilute 20 mM compound I stock solution to 25 μM with working solution, and store at 4° C. in the dark.
[0079] 2.3 Turn on the microplate reader to preheat in advance, and set its measurement parameters:...
Embodiment 2-10
[0089] Except that compound I in Example 1 was changed to compounds II-XI shown in Table 1, the same sample was measured in the same manner as in Example 1, and the obtained results are shown in Table 1.
[0090]
[0091]
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Abstract
Methods and reagent kits for determining the activity of thioredoxin reductase in a sample and the uses thereof are disclosed. The methods comprise determining the sulphydryl content of selen-compounds before and after treating the sample individually, consequently counting the activity of thioredoxin reductase in the sample by the difference of sulphydryl content before and after treating the sample. Additionally, the present invention also discloses the methods for evaluating the level of disease resistance in vivo, which comprise the step of determining the activity of thioredoxin reductase in a sample, and the step of counting the level of oxidative stress and the activity of thioredoxin reductase.
Description
[0001] This application relies on and claims the priority of Chinese patent application 200910207455.X filed on November 03, 2009, the entire contents of which are hereby incorporated by reference. Technical field: [0002] The invention belongs to the field of enzyme activity detection, in particular to a method, reagent and application for measuring the activity of thioredoxin reductase in a sample. Background technique: [0003] In my country, about 2 million people suffer from cancer every year, and 1.5 million people die of cancer, and it is still increasing, and it is gradually showing a younger trend. The direct economic loss caused by cancer to our country exceeds 100 billion yuan every year (excerpted from Beijing Morning Post 2007 / 03 / 31). According to the report of the Ministry of Health in 2007, the number of deaths due to tumors in my country has reached 1.8 million by 2006, which is 300,000 higher than that in 2005. In the early stage of tumor occurrence, the h...
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