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CuZn/Mn Superoxide Dismutase Activity Assay Kit (Hydroxylamine Method)– MSE Supplies LLC

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CuZn/Mn Superoxide Dismutase (CuZn-SOD/Mn-SOD) Activity Assay Kit (Hydroxylamine Method) - MSE Supplies LLC

CuZn/Mn Superoxide Dismutase (CuZn-SOD/Mn-SOD) Activity Assay Kit (Hydroxylamine Method)

SKU: E-BC-K022-M-96

  • £19300
  • Save £3400

CuZn/Mn Superoxide Dismutase (CuZn-SOD/Mn-SOD) Activity Assay Kit (Hydroxylamine Method)

SKU # E-BC-K022-M
Detection Instrument Microplate reader (530-570 nm, optimum wavelength: 550 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms CuZn-SOD,Mn-SOD,T-SOD
Sample Type Serum, plasma, urine, cells, cell culture supernatant, tissue
Sensitivity 1.35 U/mL
Detection Range 1.35-62 U/mL
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 70 min
Precision Average inter-assay CV: 9.600% | Average intra-assay CV: 5.100%
Other instruments required Micropipettor, Centrifuge, Incubator, Vortex mixer
Other reagents required Normal saline (0.9% NaCl)
Storage Reagent 4: -20℃, others: 2-8℃
Valid period 12 months

 

Dilution of Sample

The optimal sampling volume are different for different species, the SOD also are different for different samples. It is recommended to take 2~3 samples to do a pre-experiment, diluting a series of diluent and determine the dilution factor when the SOD inhibition ratio is 15%~55% (the optimal inhibition ratio is the range of 25%~45%) before formal experiment.

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Mouse serum 05-10
Rat serum 6-15
Urine 2-3
Human hydrothorax 2-3
10% Mouse liver tissue homogenate 100-200
10% Mouse brain tissue homogenate 20-30
10% Mouse kidney tissue homogenate 50-120
10% Rat kidney tissue homogenate 50-120
HepG2 cells (5.21mgprot/mL) 15-25
Cell culture supernatant 1

 

Note: The diluent is normal saline (0.9% NaCl).

 

Detection Principle

Superoxide anion (O2•-) produced by xanthine and xanthine oxidase system can oxidize hydroxylamine to form nitrite which appear purplish red after chromogenic reaction. The SOD in the sample has a specific inhibitory effect on superoxide anion (O2•-), can reduce the content of nitrite. The OD value is lower than control and the activity of SOD is calculated through the formula. There are two kinds of SOD (CuZn-SOD, Mn-SOD) in the cells of higher animals, and the sum of them is equal to the total SOD. The activity of Mn-SOD in the sample will lost after sample pretreatment, but the activity of CuZn-SOD will not.


Kit Components & Storage

Item Component Size 1 (48 T) Size 2 (96 T) Storage
Reagent 1 Buffer Solution 1.2 mL × 1 vial 1.2 mL × 1 vial 2-8℃, 12 months
Reagent 2 Nitrosogenic Agent 1.2 mL × 1 vial 1.2 mL × 1 vial 2-8℃, 12 months
Reagent 3 Substrate Solution 1.2 mL × 1 vial 1.2 mL × 1 vial 2-8℃, 12 months
Reagent 4 Enzyme Stock
Solution		 0.03 mL × 1 via 0.06 mL × 1 vial -20℃, 12 months
Reagent 5 Enzyme Diluent 1.2 mL × 1 vial 1.2 mL × 1 vial 2-8℃, 12 months
Reagent 6 Chromogenic
Agent A		 Powder × 1 vial Powder × 1 vial 2-8℃, 12 months
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Reagent 7 Chromogenic
Agent B		 Powder × 1 vial Powder × 1 vial 2-8℃, 12 months
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Reagent 8 Chromogenic
Agent C		 6 mL × 1 vial 6 mL × 1 vial 2-8℃, 12 months
Reagent 9 Extracting Solution 6 mL × 1 vial 12 mL × 1 vial 2-8℃, 12 months
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Microplate 96 wells No requirement
Plate Sealer 2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters Sample 1 Sample 2 Sample 3
Mean (U/mL) 2.60 29.50 48.20
%CV 5.6 4.9 4.8


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (U/mL) 2.60 29.50 48.20
%CV 10.2 9.2 9.4


Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 99%


 Sample 1 Sample 2 Sample 3
Expected Conc. (U/mL) 13 34 55
Observed Conc. (U/mL) 13.1 33.3 53.9
Recovery rate (%) 101 98 98


Sensitivity

The analytical sensitivity of the assay is 1.35 U/mL. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.