CoenzymeⅡ NADP(H) Content Assay Kit (WST colorimetry)

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辅酶Ⅱ NADP(H)含量检测试剂盒（WST显色法）

Cat:

NA0171

Assay Type:

Spectrophotometer

Brand:

sunlong medical

Specificity:

50T/24S

Storage instructions：

-20℃

Product Overview：

Components:

Acid Extract solutiont : Liquid 15 mL×1. Storage at 2-8℃.

Alkaline Extract solutiont: Liquid 15 mL×1. Storage at 2-8℃.

Reagent I : Liquid 30 mL×1. Storage at 2-8℃.

Reagent II : Powder×1. Storage at -20℃. Add 12mL of distilled water before using to dissolve it.

Reagents can be stored at 2-8°C for 4 weeks after dissolution.

Reagent III : Liquid 12 mL×1. Storage at 2-8℃.

Reagent IV: Liquid 12 mL×1. Storage at 2-8℃.

Reagent V : Liquid 70 mL×1. Storage at 2-8℃.

NADP standard: Powder×1. Storage at -20℃.Add 1.27 mL of distilled water before use to obtain a

standard of 5 µmol/mL, which can be stored at -20°C for 2 weeks.

NADPH standard: Powder×1. Storage at -20℃.Add 1.2 mL of distilled water before use to obtain a

standard of 5 µmol/mL, which can be stored at -20°C for 2 weeks.

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Unit:

Price: $185

Product PDFs

Datasheet:

Other Information
Citations
Protein Attributes
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Coenzyme II NADP(H) is widely present in animals, plants, microorganisms and cultured cells. The

determination of NADP⁺and NADPH content can calculate the content of NADP (NADPH + NADP⁺) and

the ratio of NADPH/NADP⁺, and its changes are related to the pentose phosphate pathway and

biosynthesis and Antioxidative responses are closely related. The NADPH/NADP⁺ratio is not only one of

the main markers of cellular redox state, but also plays an important regulatory role in the PPP pathway,

biosynthesis and antioxidant metabolism.

The NADP⁺and NADPH in the samples were extracted with acidic and basic extraction solutions,

respectively. Under the action of 1-mPMS, WST-1 can react with NADPH to produce water-soluble

formazan, which has a characteristic absorption peak at 450nm, while NADP⁺can be reduced to NADPH

by 6-phosphate glucose dehydrogenase, which is further detected by WST-1.

Reagents and Equipment Required but Not Provided:

Spectrophotometer, centrifuge, water bath, mortar/homogenizer, sonicator, adjustable pipette, 1 mL

glass cuvette, ice, and distilled water.

Sample Preparation：

1. Serum

Extract NADP⁺: Take 0.1 mL of serum (slurry), add 0.5 mL of acidic extract, boil for 5 minutes

(cover tightly to prevent water loss), and after cooling in an ice bath, centrifuge at 10,000g at 4°C for

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10 minutes; take 200 µL of supernatant, add an equal volume of alkaline extract ; Mix well, centrifuge

at 10,000g at 4°C for 10 min, take the supernatant, and store on ice for testing.

Extract NADPH: Take 0.1 mL of serum (slurry), add 0.5 mL of alkaline extract, boil for 5 minutes

(cover tightly to prevent water loss), after cooling in an ice bath, centrifuge at 10,000g at 4°C for 10

minutes, take 200 µL of supernatant, and add an equal volume of acidic extract ; Mix well, centrifuge

at 10000g at 4°C for 10min, take the supernatant and store on ice for testing.

2. Tissue

Extract NADP⁺: Weigh about 0.1g of tissue, add 0.5mL of acidic extract, grind in ice bath, boil for

5min (cover tightly to prevent water loss), after cooling in ice bath, centrifuge at 10,000g at 4°C for

10min, take 200µL of supernatant, add an equal volume of alkaline extract ; Mix wel, centrifuged at

10,000 g at 4°C for 10 min, and the supernatant was taken and stored on ice for testing.

Extract NADPH: Weigh about 0.1 g of tissue, add 0.5 mL of alkaline extraction solution, grind in an

ice bath, boil for 5 min (cover tightly to prevent water loss), after cooling in an ice bath, centrifuge at

10,000 g at 4°C for 10 min, take 200 µL of supernatant, and add an equal volume of acidic extract ;

Mix well, centrifuged at 10,000 g at 4°C for 10 min, and the supernatant was taken and stored on ice

for testing.

3. Bacteria or cells

Extract NADP⁺: Collect 5 million cells or bacteria, add 0.5mL of acidic extract, ultrasonically disrupt

for 1min (intensity 20% or 200W, ultrasonic for 2s, stop for 1s), boil for 5min (cover tightly to prevent

water loss), cool in ice bath, 10000g centrifuge at 4°C for 10min, take 200uL of the supernatant into

another new centrifuge tube, add an equal volume of alkaline extract to neutralize, mix well,

centrifuge at 10,000g at 4°C for 10min, take the supernatant and store it on ice for testing .

Extract NADPH: Collect 5 million cells or bacteria, add 0.5mL alkaline extract, ultrasonically

disrupt for 1min (intensity 20% or 200W, ultrasonic for 2s, stop for 1s), boil for 5min (cover tightly to

prevent water loss), cool in an ice bath, Centrifuge at 10,000g at 4°C for 10min, take 200uL of the

supernatant into another new centrifuge tube, add an equal volume of acidic extract to neutralize, mix

well, centrifuge at 10,000g at 4°C for 10min, take the supernatant, and store it on ice for testing .

Determination

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