Molecular marker for breast carcinoma diagnosis
A breast cancer, thioredoxin technology, applied in the direction of analysis materials, measuring devices, instruments, etc., can solve the problem of breast cancer correlation that has not been seen in thioredoxin
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Embodiment 1
[0026] Preparation of protein samples from normal surrounding tissues and mass tissues of breast cancer:
[0027] Breast tissue: The samples were from the First Affiliated Hospital of Kunming Medical College, Yunnan Province. All tumors have been pathologically diagnosed.
[0028] The method for preparing protein samples from normal surrounding tissues and mass tissues of breast cancer is as follows:
[0029] 1. The fresh tissue pieces removed by surgery are stored in a refrigerator at -80℃.
[0030] 2. When preparing protein, cut out tissue pieces of about 5 mm, grind them into cell pellets with a grinder on ice, and add 120 microliters of cell lysate (sodium chloride, NP-40, Tris, PMSF, aprotimin, NaN3 / 100 Ml), placed on ice for 50 minutes, then centrifuged at 12000r / min at 4°C for 15 minutes, and the supernatant was stored in a refrigerator at -80°C.
Embodiment 2
[0032] Western blot verification of thioredoxin and thioredoxin binding protein 2:
[0033] In order to confirm the differential expression of thioredoxin and thioredoxin binding protein 2, samples of the mass tissue and corresponding normal peripheral tissue proteins of 30 breast cancer patients were taken, and anti-thioredoxin and thioredoxin binding protein were used. The antibody of 2 is subjected to western blot analysis, and the specific process is briefly described as follows:
[0034] Take 7 ug total protein load for each sample, separate by 15% SDS-PAGE, transfer to the PVDF membrane (purchased from Millipore), and incubate the PVDF membrane overnight with skim milk at 4°C. Wash three times with TPBS for 5 minutes each time. The primary antibody was a mouse anti-human thioredoxin antibody (provided by Professor Yodoi Yuji, Kyoto University, Japan), incubated for 1 hour at room temperature, and washed three times with TPBS for 5 minutes each. Secondary antibody anti-mouse ...
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