Detection kit for completely and homogeneously determining glucagon and method thereof
A glucagon and detection kit technology, which is applied in the field of detection kits for completely homogeneous determination of glucagon, can solve the problems of inability to meet detection requirements, limited development and application, and low degree of automation, and achieves rapid detection. Convenience, improved accuracy and sensitivity, wide inspection results
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Embodiment 1
[0063] Embodiment 1, the preparation of glucagon calibrator
[0064] The matrix liquid of calibrator of the present invention comprises the component of following concentration:
[0065]
[0066]
[0067] Prepare the calibrator matrix solution according to the above ratio, and use this matrix solution to dilute the glucagon antigen into corresponding concentrations of calibrator, the concentrations are 1pmol / L, 4pmol / L, 16pmol / L, 32pmol / L, 128pmol / L, Add 0 pmol / L matrix solution, a total of 6 bottles, 1 mL each, and store at 2-8°C.
Embodiment 2
[0068] Example 2, Preparation of Acridan-labeled anti-glucagon antibody
[0069] Take 250μg of anti-glucagon antibody (diluted to 1mg / mL with 20mM PBS first), add 40μL of Acridan, add 710μL of 20mM PBS, and incubate on a rotator at room temperature for 1h; then use marker buffer (30mM pH 7.6 PBS buffer, 0.1w / v% sodium thiosulfate, 0.5w / v% BSA, 0.4w / v% mannitol and 0.05w / v% ProClin300) were diluted to the working concentration.
[0070] According to this method, cridan-labeled anti-glucagon antibodies with concentrations of 1.5 mg / L, 3 mg / L, 6 mg / L, 12 mg / L, 24 mg / L and 48 mg / L were prepared.
Embodiment 3
[0071] Example 3, preparation of horseradish peroxidase-labeled anti-glucagon antibody
[0072] Use a commercial HRP labeling kit to label anti-glucagon antibody, and then use label buffer (30mM pH7.6PBS buffer, 0.05w / v% VC or 0.05w / v% reduced glutathione, 0.03w / v% EC Oxyrase, 0.5w / v% mannitol, 0.5w / v% polyethylene glycol 20000, 0.1w / v% enzyme stabilizer, 0.05w / v% ProClin 300.) diluted to the working concentration.
[0073] According to this method, HRP-labeled anti-glucagon antibodies were prepared at concentrations of 0.025 mg / L, 0.05 mg / L, 0.1 mg / L, 0.2 mg / L, 0.4 mg / L, and 0.8 mg / L, respectively.
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