식물 탈수소효소 (PDHA) 활동 분석 키트
메모: 테스트 전에 예측을 위해 2~3개의 서로 다른 샘플을 채취합니다..
운영장비: Microplate Reader/ Spectrophotometer
고양이 번호: BC3125
크기:100T/48S
구성요소:
시약 I: 가루×2. 4℃에서 보관. Dissolve one bottle of powder in water to make 50 mL before use, prepare the solution when it will be use. Store at 4℃ and protect from light after prepared.
시약 II: 액체 100 mL×2. 4℃에서 보관.
시약 III: Ethyl acetate, 필수이지만 제공되지 않았습니다.
제품 설명:
The activity of plant dehydrogenase (PDHA) is largely reflecting the active state of the organism, which can directly indicate the ability of biological cells to degrade its matrix.
The hydrogen acceptor 2,3,5-triphenyl tetrazolium chloride (TTC) generates red triphenylformazone (TFF) after receiving hydrogen during cell respiration. TFF has a characteristic absorption peak at 485 nm, the PDHA activity can quantified by measuring the absorbance at 485 nm.
필요하지만 제공되지 않는 시약 및 장비:
Microplate Reader or spectrophotometer, 욕조, 책상 원심분리기, 욕조, 피펫, 마이크로 유리 큐벳/96웰 평면 바닥 플레이트 (non-polystyrene/polypropylene material), 모르타르/균질화기, ethyl acetate (express delivery is not allowed), 얼음과 증류수.
절차:
나. Complex 추출:
모으다 0.1 g의 조직, 씻다 3 또는 4 times with double steam water, blot dry with filter paper and set aside.
II. 결정 절차:
- Preheat spectrophotometer/ microplate reader for 30 분, 파장을 조정하여 485 nm, set zero with ethyl acetate.
- 다음 시약을 추가하십시오 5 mL EP tubes:
| 시약 | Contrast tube (AC) | 시험관 (에) |
| 견본 (g) | 0.1 | 0.1 |
| 시약 I (밀리리터) | – | 1 |
| 시약 II (밀리리터) | 2 | 1 |
| Mix thoroughly and stand in dark for 3 hours at 37℃, ice bath for 5 minutes immediately after take out. Discard the filtrate, blot dry the sample with filter paper, place in mortar / homogenizer. | ||
| 시약 III (밀리리터) | 1 | 1 |
III. 계산:
ㅏ. micro glass cuvette (light path of the cuvette, 1 센티미터)
단위 정의: One unit of enzyme activity is defined as the amount of enzyme increases the absorbance of every 0.01 for per hour every mg tissue protein in the reaction system.
PDHA Activity(U/g/h) =ΔA÷0.01÷T÷W=33×ΔA÷W
비. 96 우물 접시 (light path of the cuvette, 0.6 센티미터)
단위 정의: One unit of enzyme activity is defined as the amount of 1 mg of tissue increases the absorbance of every 0.005 for per hour in the reaction system.
PDHA Activity(U/g/h) =ΔA÷0.005÷T÷W=66.7×ΔA÷W
티: 반응 시간 (시간), 3 시간; 여: 샘플 중량, g.
메모
- After prepared, Reagent I should store at 4℃, protect from light and used within one week. If it turns red, it cannot be used.
- Reagent III is volatile and toxic. For your health, please wear lab clothes, masks, and latex gloves.
- Ice bath to stop the reaction immediately after the dark reaction was completed. Discard the filtrate, blot dry the sample with filter paper as much as possible.
- 테스트 전에 예측을 위해 2~3개의 서로 다른 샘플을 채취합니다.. Dilute the supernatant if the absorbance is higher, 공식에서 희석 시간을 곱하십시오.
- If test with a 96 바닥이 평평한 접시, polystyrene, or polypropylene material 96 well flat-bottom plate is not recommended.
실험예:
- Weigh 1g aloe leaves, wash them with double distilled water for 3–4 times, absorb the water with filter paper, 결정 단계에 따라 작동, measure and calculate with 96 우물 접시, ΔA=AT-AC= 0.244-0.146 = 0.098, calculate the enzyme activity.
Dehydrogenase activity (U/g 질량) = 66.7×ΔA÷W = 6.5366 U/g 질량.
관련 상품：
BC2030/BC2035 Isocitrate Lyase (ICL) 활동 분석 키트
BC3170/BC3175 Acetokinase (ACK) 활동 분석 키트
BC2010/BC2015 Glycollic Oxidase Activity Assay Kit
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