Detection kit and detection method for soluble urokinase-type plasminogen activator receptor
A detection kit, plasminogen technology, applied in measurement devices, biological material analysis, instruments, etc., can solve the problems of high specificity, low detection sensitivity and accuracy of ELISA, and improve detection sensitivity and specificity. performance, optimized response time, simple operation
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[0024] 1. Preparation of Magnetic Separation Reagents
[0025] 1. Preparation of magnetic particle buffer
[0026](1) Weigh TRIS 4.58g / L, NaCl 6.81g / L, TRITON X-100 0.5mL / L in a 1L container; (2) Measure Proclin-300 0.5-1mL in a 1L container; measure 800mL for purification Put water in the above container and stir thoroughly; (3) adjust the pH value to 7.95-8.05; (4) weigh 3-5g of BSA into the above 1L container, and wait until it is fully dissolved; (5) finally make it to 1000mL, 0.22 um microporous filter; mark it well and store it in a cold storage at 2-8°C for later use.
[0027] 2. Preparation of magnetic separation reagents
[0028] (1) Dissolve 1.0 mg disuccinimidyl suberate in 50 μL DMSO to obtain a disuccinimidyl suberate solution; dissolve 2 mg streptavidin in 0.1 mol / 1 mL of phosphate buffered saline to a total volume of 1 mL to obtain a streptavidin solution;
[0029] (2) Determine the input amount of disuccinimidyl suberate, draw the disuccinimidyl suberate s...
Embodiment 1
[0070] This embodiment provides a soluble urokinase-type plasminogen activator receptor detection kit, including: magnetic separation reagents, antibody reagents, calibrator, quality control products, concentrated cleaning solution and substrate solution; wherein, the antibody reagent Containing alkaline phosphatase-labeled antibody and biotin-labeled antibody, the magnetic separation reagent includes magnetic particles coated with streptavidin.
[0071] 1. Preparation of Magnetic Separation Reagents
[0072] 1. Preparation of magnetic particle buffer
[0073] (1) Weigh TRIS 4.58g / L, NaCl 6.81g / L, TRITON X-100 0.5mL / L in a 1L container; (2) Measure Proclin-300 0.5mL in a 1L container; measure 800mL purified water (3) Adjust the pH value to 8.00; (4) Weigh 4g of BSA into the above 1L container and wait for it to fully dissolve; (5) Finally, set the volume to 1000mL and filter through a 0.22um microporous filter ; Mark it and store it in a cold room at 2-8°C for later use.
...
Embodiment 2
[0105] This embodiment provides a method for detecting soluble urokinase-type plasminogen activator receptor (suPAR) using a soluble urokinase-type plasminogen activator receptor detection kit, comprising steps:
[0106] S1: Add 50 μL standard substance, 50 μL quality control substance and 50 μL serum of the sample to be tested to the bottom of the chemiluminescence reaction tube;
[0107] S2: Add 50 μL of antibody reagent to the bottom of the chemiluminescence reaction tube;
[0108] S3: Gently shake the reaction tube for 30 seconds with a multi-functional homogeneous mixer, and then place it in a thermostat at 37°C for 15 minutes;
[0109] S4: Add 50 μL of magnetic separation reagent to the bottom of the chemiluminescence reaction tube;
[0110] S5: Repeat step S3 once;
[0111] S6: Put the reaction tubes on the magnetic separator, make sure that the bottom of each tube is in contact with the surface of the separator, and settle for 2 minutes; then slowly invert the separa...
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