Application and kit of combined markers of igf2bp3 and afp in preparation of liver cancer diagnosis and treatment evaluation kit
A diagnostic kit and marker technology, applied in biological tests, instruments, measuring devices, etc., can solve problems such as false negatives, achieve broad prospects and application value, significant diagnostic significance, and avoid false positive and false negative results.
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Embodiment 1
[0047] The IGF2BP3 detection kit provided in this embodiment includes the first antibody for specifically binding to IGF2BP3. The primary antibody is a mouse anti-human IGF2BP3 monoclonal antibody. Wherein, the amino acid sequence of IGF2BP3 is shown in SEQ ID NO.1.
[0048] The method for detecting the concentration of IGF2BP3 in the sample to be tested using the IGF2BP3 detection kit provided in the examples in this embodiment is as follows:
[0049] S1: Coating primary antibody. Add 50 μL of pH 9.6 carbonate buffer solution and 100 μL of 5 μg / mL mouse anti-human IGF2BP3 monoclonal antibody to each well of a microtiter plate (purchased from Greiner, Cat. No. 705071), and coat overnight at 4°C. , each well was washed 3 times with 300 μL of washing solution, soaking for 2 minutes each time (hereinafter referred to as plate washing).
[0050] S2: Close the excess gap. Add 100 μL bovine serum albumin (BSA) with a mass fraction of 2%, block at 25° C. for 2 hours, and wash the...
Embodiment 2
[0058] The IGF2BP3 detection kit provided in this example is roughly the same as the IGF2BP3 detection kit provided in Example 1, the difference is that the IGF2BP3 detection kit in this example also includes an enzyme-labeled Secondary antibody. The second antibody is rabbit anti-mouse anti-human IGF2BP3 polyclonal antibody, and the enzyme is horseradish peroxidase. The second antibody is stored in a stabilizer, which includes PBS with a volume fraction of 68%, BSA with a mass fraction of 30% and NaN with a mass fraction of 0.02% 3 .
[0059] The detection method of using the IGF2BP3 detection kit provided in this implementation to detect the concentration of IGF2BP3 in the sample to be tested is the same as that in Example 1.
Embodiment 3
[0061] The IGF2BP3 detection kit provided in this example is roughly the same as the IGF2BP3 detection kit provided in Example 2, except that the stabilizer in this example includes PBS with a volume fraction of 66% and a mass fraction of 28% BSA and NaN with a mass fraction of 0.018% 3. In addition, the IGF2BP3 detection kit in this embodiment also includes a solid phase carrier and a blocking membrane, and the solid phase carrier is a 96-well microtiter plate (12 strips×8 wells).
[0062] The detection method of using the IGF2BP3 detection kit provided in this implementation to detect the concentration of IGF2BP3 in the sample to be tested is the same as that in Example 1.
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