Solarbio 총 항산화 능력 (T-AOC) 분석 키트

총 항산화 능력 (T-AOC) 분석 키트

메모: 테스트 전에 예측을 위해 2~3개의 서로 다른 샘플을 채취합니다..

운영장비: 분광 광도계

고양이 번호: BC1310

크기：50T/48S

구성요소:

용액 추출: 액체 50 밀리리터×1. 4℃에서 보관, precool before use.

시약 I: 액체 35 밀리리터×1. 4℃에서 보관.

시약 II: 액체 20 밀리리터×1. Storage at 4℃ in shadow.

시약 III: 액체 5 밀리리터×1. Storage at 4℃ in shadow.

기준: 가루×1, 10 mg of FeSO4·7H2O. 작업 솔루션: 추가하다 0.9 mL of distilled water and 20μL of concentrated sulfuric acid (H2SO4) to forms 40 µmol/mL FeSO4 standard solution. Solution mixture (prepare when the solution will be used): 시약 I : 시약 II: Reagent III= 7:1:1, incubate at 37℃ before use.

제품 설명:

This kit is used to detect the total antioxidant levels of antioxidants and antioxidant enzymes in the samples. It is mainly used in the study of biological, medical and pharmaceutical studies to detect the total antioxidant capacity of antioxidant solutions.

In acid environment, Fe3+ -TPTZ are reduced to blue Fe2+ -TPTZ. The color reaction reflects the total antioxidant capacity.

필요하지만 제공되지 않는 시약 및 장비:

분광 광도계, 항온수욕, 저온 원심분리기, 1 mL glass cuvette and distilled water.

절차:

나. 샘플 준비:

1. 혈청, 혈장, 타액, or urine samples

혈장 (anticoagulation with heparin or sodium citrate, avoid using EDTA), 원심분리기 5000 rpm/min for 10 분, take supernatant for test. Take serum, saliva or urine samples for direct determination. 또한, you can store at -80℃ and detect within 30 날.

2. Cells or tissue 견본

가져가다 1-2 million cells or 0.1 g의 조직, 추가하다 1.0 mL의 추출 용액. Use homogenate or ultrasound to fully break up cells and release antioxidant, 원심분리기 10000 r/min and 4℃ for 5 분, take supernatant for test. Measure the concentration of protein if needed.

II. Determination procedure:

1. 묽게 한 40 μmol/mL FeSO4 standard solution to 0.1, 0.05, 0.025, 0.0125, 0.00625, 0.003125 μmol/mL, 가져가다 500 μL of standard solution (distilled water for blank control), add to 500 시약 II의 μL. Mix thoroughly for 10 분, detect the absorbance in 593 nm, calculate ΔA=AS-AB. (처럼: standard solution tube, AB: blank control tube.) The final concentration of Fe2+ is 0.05、0.025、0.0125、0.00625、

0.003125、0.00156 μmol/mL.

2. Preheat the spectrophotometer 30 분, 파장을 조정하다 593 nm and set zero with distilled
3. 다음 목록으로 시약을 추가하세요.:

III. 계산:

1. Create standard curve

Take the Fe2+ final concentration as X-axis, △A as Y-axis, create standard curve, get linear regression equation y=kx+ b, take ΔA’ into the equation to get x (μmol/mL).

2. 단위 정의: the sample antioxidant capacity is indicated by the standard liquid ion concentration required for the same absorbance change(ΔA).

ㅏ. Protein 집중:

총 항산화 능력 (μmol/mg prot) = x × Vrv÷ (Vs× Cpr) = 34× x ÷ Cpr

비. 견본 weight

총 항산화 능력 (μmol/g weight) = x × Vrv÷ (Vs ÷ Vsv ×W) =34× x÷ W

씨. Cell amount

총 항산화 능력 (μmol/104cell) = x × Vrv÷ (Vs ×Vsv÷n) = 34× x÷ n

디. Solution 용량

총 항산화 능력 (μmol/mL) =x× Vrv÷ Vs =34×x

로프: total reaction volume, 1.02 밀리리터; 대: 샘플량, 0.03 밀리리터;

VSV: 추출량, 1 밀리리터; 여: 샘플 중량, g;

심폐소생술: 샘플 단백질 농도, mg/mL;

n: cell amount, unit based on 104 (ten thousand).

메모:

1. Reagent II is irritated to human body, please wear lab clothes and latex
2. The samples should not be appeared blue under acidic condition, or it will interference sample result of the
3. Detergent such as Tween, Triton, NP-40 and reductants such as DTT, mercapto ethanol should not be added in the
4. If the absorbance value determined by the sample is beyond the standard curve range, the sample should be diluted or concentrated properly before
5. The kit should be store at2-8℃.

Examples:

1. Add 0.1g shamrock to 1mL extract solution and grind thoroughly on ice, take supernatant, follow the determination procedure to operate, with 96-well flat-bottom plates to calculate: ΔA=A(티)-ㅏ(비)=0.909-0.148=0.761, standard curve: y=21.056x-0.0087, calculate x=0.037, according to mass of sample to calculate Total antioxidant capacity (μmol/g mass) =34×x÷W=34×0.037÷0.1=12.85 μmol/g mass.

참고자료：

[1] Pellegrini N, Serafini M, Salvatore S, 외. Total antioxidant capacity of spices, dried fruits, nuts,

pulses, cereals, and sweets consumed in Italy assessed by three different in vitro assays[제이]. Molecular nutrition & food research, 2006, 50(11): 1030-1038.

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