Application of protein muc13 in preparation of reagents for diagnosing intrahepatic cholangiocarcinoma
A technology for intrahepatic cholangiocarcinoma and protein, applied in the field of medicine, to achieve highly specific effects
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Embodiment 1
[0046] Example 1 Determination of Candidate Serum Biomarkers
[0047] The inventors of the present invention used the advantages of high-throughput and rapid analysis of human proteome chips to analyze 60 serum samples related to intrahepatic cholangiocarcinoma (30 serum samples from patients with intrahepatic cholangiocarcinoma and 30 serum samples from healthy people). Comparing the difference between samples from patients with intrahepatic cholangiocarcinoma and healthy people, it was found that the expression level of MUC13 protein in patients with intrahepatic cholangiocarcinoma was significantly higher than that in healthy people, suggesting that MUC13 protein can be used as a candidate serum for intrahepatic cholangiocarcinoma Biomarkers for early diagnosis and effective treatment of intrahepatic cholangiocarcinoma.
Embodiment 2
[0048] Example 2 Application of MUC13 Protein in Diagnosis of Intrahepatic Cholangiocarcinoma
[0049] 1. Expression, purification and identification of MUC13:
[0050] The protein MUC13 is obtained from genetically engineered Saccharomyces cerevisiae, induced overexpression by galactose, separated and purified by agarose affinity medium (glutathione), and the results of silver staining quantification and Western-Blotting identification were respectively Such as figure 1 shown.
[0051] 2. Preparation of serum samples:
[0052] After all the specimens were left at room temperature for 2 hours, centrifuge at 2000g for about 5 minutes, and the supernatant was taken for immediate detection; or aliquoted, and the specimens were stored at -20°C or -80°C, but repeated freezing and thawing should be avoided. Thawed samples should be centrifuged again prior to testing. The tested samples cannot contain NaN 3 , because NaN 3 Inhibits horseradish peroxidase (HRP) activity.
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