Application of ZNF74 protein as gastric cancer serum biomarker and kit thereof
A technology of serum markers and biomarkers, applied in the field of biological sciences, to achieve high specificity and simple operation
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[0037] 1. Expression, purification and identification of ZNF74 protein:
[0038] The human protein ZNF74 is obtained by using conventional methods to induce over-expression with galactose from genetically engineered Saccharomyces cerevisiae, and then separated and purified by glutathione on agarose affinity medium, and identified by Western-Blotting.
[0039] 2. Preparation of serum samples:
[0040] Whole blood samples were left at room temperature for 2 hours, then centrifuged for 20 minutes, and the supernatant was collected for aliquots and stored at -80°C. Thawed serum samples need to be centrifuged again before they can be used for detection.
[0041] 3. Preparation methods of various buffers and reagents for ELISA method:
[0042] (1) Coating buffer: 0.05M sodium carbonate-sodium bicarbonate (pH 9.6)
[0043]
[0044] (2) Sample diluent: PBS solution with pH 7.4
[0045]
[0046](3) Washing solution: PBST solution with pH 7.4
[0047]
[0048] (4) Blocking...
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