Proline aminopeptidase detection reagent, reaction pad, preparation method and kit thereof
A proline aminopeptidase and detection reagent technology, which is applied in biochemical equipment and methods, biological testing, measuring devices, etc., can solve problems such as lack of accuracy, and achieve the effects of good accuracy, high sensitivity, and accurate detection
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[0044] The preparation method of the reaction pad includes the following steps:
[0045] 1) Material preparation: prepare dithiothreitol, sucrose, polyvinylpyrrolidone, Z-glycyl-proline-p-nitroaniline and bovine serum albumin according to the above weight parts;
[0046] 2) Prepare buffer: prepare 0.01mol / L~1mol / L Tris-HCl buffer;
[0047] 3) preparing soaking solution: dissolving dithiothreitol, sucrose, polyvinylpyrrolidone, Z-glycyl-proline-p-nitroaniline and bovine serum albumin in Tris-HCl buffer to obtain soaking solution;
[0048] 4) soaking solution treatment: the filter paper is fully soaked in the soaking solution for 10-20s, then taken out, and then dried at a temperature of 20-75°C and a time of 30-40min to obtain a reaction pad.
[0049] More specifically, the filter paper can be Whatman glass fiber filter paper, GF / A Whatman's fiber filter paper Grade 3 or ordinary quantitative filter paper.
[0050] Use the reaction pad to prepare the kit: place the reaction p...
Embodiment 1
[0052] Examples 1 to 3 disclose proline aminopeptidase detection reagents. Three groups of raw material components are prepared according to Table 1, and Tris-HCl buffers with different concentrations are prepared according to Table 2; group A is used in Example 1, and group B is used. For Example 2, Group C was used for Example 3.
[0053] Dithiothreitol, sucrose, polyvinylpyrrolidone, Z-glycyl-proline-p-nitroaniline and bovine serum albumin were dissolved in 100 mL of Tris-HCl buffer in turn to obtain proline aminopeptidase detection reagents.
[0054] Table 1 Composition and content of detection reagents
[0055]
[0056] Table 2 Preparation parameters of Tris-HCl buffer
[0057]
[0058]Detect the detection reagents obtained in Examples 1 to 3: prepare proline aminopeptidase standard solution with different enzyme activity concentrations respectively, the proline aminopeptidase standard solution uses physiological saline as solvent, and detect Example 1 ~3 sensiti...
Embodiment 4~6
[0065] Embodiment 4~6 discloses a kind of reaction pad and preparation method thereof:
[0066] 1) Prepare materials according to Table 1, prepare Tris-Hcl buffer solution according to Table 2; Group A is used in Example 4, Group B is used in Example 5, and Group C is used in Example 6;
[0067] 2) Prepare soaking solution: dissolve dithiothreitol, sucrose, polyvinylpyrrolidone, Z-glycyl-proline-p-nitroaniline and bovine serum albumin in Tris-Hcl buffer to obtain soaking solution;
[0068] 3) Soaking solution treatment: put the filter paper in the soaking solution and fully soak it for 10-20 seconds, then take it out, and then dry it at a temperature of 45° C. for 40 minutes to obtain a reaction pad.
[0069] Detect the reaction pads obtained in Examples 4 to 6: prepare standard proline aminopeptidase solutions with different enzyme activity concentrations respectively, and use physiological saline as a solvent for the proline aminopeptidase standard solution, and test Example...
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