Kit for determining ornithine decarboxylase by using marker immunological technique and its determination method
A technology of ornithine decarboxylase and kit, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of radioactive pollution, low sensitivity, complicated operation, etc., and achieve simple operation, no pollution, and high sensitivity Effect
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Embodiment 1
[0070] Detect ornithine decarboxylase in tissue extracts, plasma, serum or other tissue fluids. Take the required amount of microwell strips coated with anti-ornithine decarboxylase antibody, add 50ul tissue extract, and make standard 0, 10, 20, 40, 100 ng / ml and negative control at the same time, add biological to each well 50ul of labeled anti-ornithine decarboxylase antibody, incubate at 37°C for 1 hour, wash 5 times, add 100ul horseradish peroxidase-labeled streptavidin to each well, incubate at 37°C for 40 minutes, wash , Add 50ul each of the developer A and B to each well. After avoiding light at 37°C for 10-15 minutes, add 50ul stop solution to each well, and test at 450nm on the microplate reader. Draw a standard curve with the OD value of each standard well and the corresponding concentration, and check the corresponding concentration from the standard curve according to the OD value of the sample well.
Embodiment 2
[0072] Determination of ornithine decarboxylase in cells (drug sensitivity test). Take 4ml of the patient’s venous blood, use heparin to prevent coagulation, separate lymphocytes with lymphocyte separation solution, and then count and adjust the number of cells to 107 / ml, 100ul per hole, and contain 2ug / ml PHA and different concentrations of anti-cancer drugs such as: Cyclophosphamide 0,10 -3 , 10 -7 , 10 -9 , Vincristine 0,10 -5 , 10 -7 , 10 -9 , Make a control hole at the same time, put 5% CO 2 Incubate in an incubator at 37°C for 18-24 hours, centrifuge and wash 3 times, then routinely fix and block, add 100ul biotin-labeled anti-ornithine decarboxylase antibody to each well, 37°C for 1-3 hours, wash 5 times , Add 100ul of horseradish peroxidase-labeled avidin to each well, wash 5 times at 37℃ for 30-60 minutes, add 50ul each of chromogenic reagent A and B to each well, 37℃ for 10-15 minutes, microplate reader The OD value is measured at 450nm, and the OD value can be used to d...
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