Method for detecting catalase activity in human serum
A catalase and human detection technology, which is applied in the field of biology, can solve the problems of narrow detection range, low sensitivity, and low stability, and achieve the effects of strong sensitivity, wide detection range, and accurate results
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Embodiment 1
[0024] A method for detecting catalase activity in human serum, using catalytic fluorescence spectrometry, using H 2 O 2 -OPD-Fe 2+ Fluorescence release system to detect catalase activity in human serum.
[0025] First, obtain serum samples of healthy men aged 11 years from the physical examination center without other processing;
[0026] Next, use a micro sampler to accurately pipette 5 μL of serum sample and 1.20 mL of H with a concentration of 1 mmol / L. 2 O 2 Mixing, adding 1.00mL of o-phenylenediamine with a concentration of 1mmol / L, 1.00mL of a ferrous sulfate solution with a concentration of 1mmol / L, and 5.00mL of an acetate solution with a concentration of 0.1mol / L, followed by the reaction at room temperature for 10min. The pH value of the salt solution was 4, and the reaction was allowed to stand for 1 min. After adding 1.00 mL of TritonX-100 with a concentration of 1 mmol / L, the volume was diluted to 50 mL. Measure the fluorescence intensity F;
[0027] Again,...
Embodiment 2
[0030] Adopt the serum sample of the healthy male who is 11 years old in embodiment 1, do not do other processing;
[0031] First, use a micro sampler to accurately pipette 5 μL of serum sample and 1.20 mL of H with a concentration of 1 mmol / L. 2 O 2 Mixing, after 30min reaction at room temperature, add 1.00mL of o-phenylenediamine with a concentration of 1mmol / L successively, 1.00mL of a ferrous sulfate solution with a concentration of 1mmol / L, and 5.00mL of an acetate solution with a concentration of 0.1mol / L, wherein ethyl acetate The pH value of the salt solution was 4, and the reaction was allowed to stand for 5 minutes. After adding 1.00 mL of TritonX-100 with a concentration of 1 mmol / L, the volume was diluted to 50 mL. Measure the fluorescence intensity F;
[0032] Next, take 5 μL of fresh serum sample, do the enzyme boiling treatment, repeat the above steps, and measure the fluorescence intensity F of the enzyme blank system 0 ;
[0033] Finally, calculate the enz...
Embodiment 3
[0035] Adopt the serum sample of the healthy male who is 11 years old in embodiment 1, do not do other processing;
[0036] First, use a micro sampler to accurately pipette 5 μL of serum sample and 1.20 mL of H with a concentration of 1 mmol / L. 2 O 2 Mixing, adding 1.00mL of o-phenylenediamine with a concentration of 1mmol / L, 1.00mL of a ferrous sulfate solution with a concentration of 1mmol / L, and 5.00mL of an acetate solution with a concentration of 0.1mol / L, followed by the reaction at room temperature for 20min. The pH value of the salt solution was 4, and the reaction was allowed to stand for 3 min. After adding 1.00 mL of TritonX-100 with a concentration of 1 mmol / L, the volume was diluted to 50 mL, and the emission wavelength of the 930N fluorophotometer was selected as 510 nm. Measure the fluorescence intensity F;
[0037] Next, take 5 μL of fresh serum sample, do the enzyme boiling treatment, repeat the above steps, and measure the fluorescence intensity F of the en...
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