Superoxide Dismutase (SOD) Assay Kit (WST-1 method)

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Product name: Superoxide Dismutase (SOD) Assay Kit (WST-1 method)


Catalog number: E-BC-K020


Size: 48T/96T

Detection method: Colorimetric method

Detection instrument: Microplate reader

 

Valid period: 3 months 


Lead Time: 7~10 days 

 

 

Experimental instrument

96-well microplate, Micropipettor, 37℃ incubator, Microplate reader (450 nm)

 

Application

This kit adopts the WST-1 to measure SOD activity. It can used to measure SOD activity of serum, plasma, animal tissue, plant tissue, culture cells, whole blood, erythrocyte, etc.

 

Detection significance

Superoxide dismutase plays a crucial role in the balance between oxidation and antioxidation in body. SOD can clean superoxide anion free radicals to protect cells from damage. The assay kit adopts xanthine oxidase method (hydroxylamine method) to determinate SOD activity. This method can determinate SOD activity in serum (plasma), cerebrospinal fluid, pleural effusion, ascites, renal dialysis fluid, urine, semen, red blood cell, white blood cell, platelet, myocardial cells, tumor cell culture, kinds of animals and plant tissues and cells and subcellular levels (mitochondria and microsome), and in microbial, drugs, food, beverage, cosmetics. Hydroxylamine method not only can determinate the total SOD activity, but also Mn-SOD and Cu, Zn-SOD activity.

 

Operation steps

1.  Sample pre-treatment

(1) Serum (Plasma): Centrifuge the serum (plasma) at 3500 rpm for 10 min if it’s turbid, take the supernatant to measure. The clarified serum (plasma) was diluted into different concentrations with normal saline to do a pre-experiment.

(2)  Tissue (animal) sample: Weigh the tissue accurately. Adding 9 times of the volume of phosphate buffer: (0.1mol/L, pH 7-7.4) according to the proportion of weight (g): Volume (mL) =1:9. Homogenized mechanically with a homogenizer under ice-bath condition, then centrifuge at 2500r / min for 10 min. Take the supernatant to determine.

(3)  Tissue (plant) sample: Weigh the tissue accurately. Add 9 times of the volume of phosphate buffer: (0.1mol/L, pH7-7.4) according to the proportion of weight (g): Volume (mL) =1:9. Homogenized mechanically with a homogenizer under ice-bath condition, then centrifuge at 3500 rpm for 10 min. The supernatant was diluted into different concentrations with phosphate buffer to do a pre-experiment.

 

2.  Operation table

Components

Control well

Blank control well

Sample well

Sample blank well

Sample (μL) 

 

 

20

20

ddH2(μL) 

20

20

 

 

Enzyme working solution (μL) 

20

 

20

 

Dilution buffer (μL) 

 

20

 

20

Substrate working solution (μL) 

200

200

200

200

Mixing, incubation for 20 min at 37, and measuring spectrophotomete at a wavelength of 450 nm.

[Notices]: Set 1~2 wells of Control, Blank control, Sample blank for each experiment.


Product performance index

1.  Stability: intra-assay CV = 5.05%, inter-assay CV = 3.32%.

2.  Detection Limit: 0.5 U/mL.

3.  ODControl≥0.2

4.  Reaction temperature: 37

 


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