Quantitative determination method for saccharified protein
A technology for the quantitative determination of glycated protein, which is applied in the field of medical testing, can solve problems such as prolonged measurement time, increased background, and increased measurement error, and achieves the effects of convenient sampling, less pain for patients, and an objective percentage of glycated protein
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Embodiment 1
[0083] Example 1, the determination of glycated hemoglobin.
[0084] · Preparation of solid-phase antibodies:
[0085] Antibody A, mouse anti-human hemoglobin monoclonal antibody, American Abcam Company, model: AB401 Antibody A needs to be coated on a microwell plate (Sigma-Aldrich, M0156), and the antibody concentration is between 1.0 μg / ml and 10.0 μg / ml Between, coating buffer, 0.1M sodium bicarbonate buffer, pH 9.6. Coating volume: 200 microliters / microwell, overnight at room temperature. Then, 300 μl / well of blocking buffer (Sigma-Aldrich, C9483) was used to block overnight at room temperature. After drying, dry and store for later use.
[0086] · Preparation of antibody-labeled enzyme conjugate
[0087] Antibody B, mouse anti-human glycosylated hemoglobin monoclonal antibody, American Abcam Company, model: AB47149 Antibody C, mouse anti-human hemoglobin monoclonal antibody, American Abcam Company, model: AB402
[0088] Antibody B and Antibody C were each conjugated w...
Embodiment 2
[0132] Example 2, Determination of glycated albumin.
[0133] · Preparation of solid-phase antibodies:
[0134] Antibody A, goat anti-human serum albumin polyclonal antibody, American Abcam Company, model: AB19180, antibody A was coated on a microwell plate (Sigma-Aldrich, MO156), and the preparation method was the same as in Example 1.
[0135] · Preparation of antibody-labeled enzyme conjugate
[0136] Antibody B, mouse anti-human serum glycosylated albumin monoclonal antibody, American USBiological Company, model: A1327-55; Antibody C: mouse anti-human serum albumin monoclonal antibody, American Abcam Company, model: AB7793
[0137] Antibody B and Antibody C need to be conjugated with horseradish peroxidase (Sigma-Aldrich, P8415), respectively. Become an antibody-labeled enzyme conjugate. Conjugation method The preparation method is the same as in Example 1.
[0138] · Preparation of human serum albumin and human serum glycated albumin standards:
[0139] Human serum a...
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