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Total Superoxide Dismutase (T-SOD) Activity Assay Kit (Hydroxylamine Method)

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Catalog number:E-BC-K019-S

Size:

100Assays
50Assays

Qty:

- +

Price: $260

Detection method: Colorimetric method

Detection instrument: Spectrophotometer

Valid period: 6 months

Lead Time: 7~10 daysWelcome to order from local distributors.

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Datasheet
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General information

Detection significance

According to the literature, superoxide dismutase exists in all oxygen-metabolizing cells to protect cells from excessive superoxide. Under the action of SOD, two superoxide anions were converted to oxygen and hydrogen peroxide.

In mammals, there are three different forms of SOD: CuZn-SOD, Mn-SOD and EC-SOD (an extracellular form of SOD). Cu-Zn SOD exists in the cytoplasmic and mitochondrial membrane spaces of the cells, while Mn-SOD is located in the mitochondrial matrix.

Detection principle

The superoxide anion free radical (O₂^-) can be produced by xanthine and xanthine oxidase reaction system，O₂^- oxidize hydroxylamine to form nitrite, it turn to purple under the reaction of developer. When the measured samples containing SOD, the SOD can specifically inhibit superoxide anion free radical (O₂^-). The inhibitory effect of SOD can reduce the formation of nitrite, the absorbance value of sample tube is lower than control tube. Calculate the SOD of sample according to the computational formula.

The key point

1. Determine optimal sampling volume of each sample before formal experiment. Calculate the inhibition ratio of serial sampling volume, and choose the optimal sampling volume when inhibition ratio in the range of 25%~45%.

2. The optimal sampling volume are different for different species, the SOD also are different for different samples. So it is best to do a pre-test to determining optimal sampling volume for a new sample.

3. It is best to reserve 3 paralleled tubes with different sampling volumes in pre-test for determining the optimal sampling volume. The sampling volume in examples as median, increase by 10 μL and decrease by 10 μL. Take the pre-test with 3 paralleled tubes and 1 control tube to determining the optimal sampling volume.

4. Adjust sampling volume: If inhibition ratio > 55%, need to dilute the sample or decrease the sampling volume than take the test. If inhibition ratio < 15%, need to increase the sampling volume.

5. All the reagents should be prepared at the day before the experiment, in order to let the reagents dissolve fully. Please bring all the reagents and samples to room temperature for 30 min before the assay.

6. The incubation time is 40 min, the incubation time can be extended to 45 min when the room temperature is lower than 20℃. Ensure the incubation temperature is 37℃.

7. EDTA should not be as anticoagulation, suggest to use heparin plasma.

Operation procedures

Operation steps

1. Sample tube: add 1 mL of Reagent 1 working solution and a* mL sample to the Sample tubes.

Control tube: add 1 mL of Reagent 1 working solution and a* mL double-distilled water to the Controltubes.

2. Add 0.1 mL of Reagent 2, 0.1 mL of Reagent3, 0.1 mL of Reagent 4 working solution successively into the tubes of Step 1.

3. Mix fully with a vortex mixer, incubate for 40 min at 37 ℃.

4. Add 2 mL of Chromogenic agent into the tubes of Step 3.

5. Mix fully and stand for 10 min at room temperature.

6. Set to zero with double-distilled water and measure the OD value of each tube at 550 nm with 1 cm optical path quartz cuvette.

[Note]: If the optimal sampling volume (a*) is the same, only one control tube need to be assay.

Operation table

Reagent	Sample tube	Control tube
Reagent 1 working solution (mL)	1.0	1.0
Sample (mL)	a*
Double distilled water(mL)		a*
Reagent 2 (mL)	0.1	0.1
Reagent 3 (mL)	0.1	0.1
Reagent 4 working solution (mL)	0.1	0.1
Mix fully with a vortex instrument, incubate for 40 min at 37 ℃.
Chromogenic agent	2	2
Mix fully and stand for 10 min at room temperature. Set to zero with double-distilled water and measure the OD value of each tube at 550 nm with 1 cm diameter cuvette.

Performance characteristics

Technical parameter

Detection range	4.7-166 U/mL	Average inter-assay CV	6.3%
Sensitivity	4.7 U/mL	Average intra-assay CV	2.8%
Average recovery rate	105%

Show all (3)
Reviews (3)
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Verified Customer

M*sSubmitted [ Jun 12 2019 ]

Application:Biochemical
Description:The T-SOD activity in HepG2 cell supernatant was detected by the SOD kit, By asking the loading quantity of technical support samples, they gave a reference loading quantity and the loading amount of the formal experiment is similar to that of the reference. At this point, it\'s a trusty assay kit

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Verified Customer

M**kSubmitted [ May 29 2019 ]

Application:Biochemical
Description:The instructions tell us the treatment method of samples in detail,But he preparation part of this biochemical Assay kit is a little too much.

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Verified Customer

r****eSubmitted [ May 29 2019 ]

Application:Biochemical
Description:I have consulted many literatures and know that the results of SOD activity detection by different methods will be different, so it is more difficult to find the literature of SOD activity range. I think the best part of this kit is the ‘Reference values for samples’ section in the instruction, which gives the reference range of SOD activity of various samples. Using this kit (E-BC-K019) to detect human plasma samples, the results are within the reference range provided by the kit, which saves me a lot of work.

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