Superoxide dismutase (SOD) ELISA test kit works - Database & Sql Blog Articles

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Kaixin micro test

Test probe P100-M3

Experimental Principle

The kit is based on a double-antibody one-step sandwich ELISA method. The microwells are pre-coated with superoxide dismutase (SOD) capture antibodies. After adding the samples, standards, and HRP-labeled detection antibodies in sequence, the mixture is incubated and washed thoroughly. A TMB substrate is then used to develop the color. Under the catalysis of peroxidase, TMB turns blue and finally becomes yellow when an acid is added. The intensity of the color is directly proportional to the SOD concentration in the sample. The absorbance at 450 nm is measured using a microplate reader, and the sample concentration is calculated accordingly.

Kit Composition

Result Calculation

To calculate the results, plot the OD values of the standard solutions on the x-axis and their corresponding concentrations on the y-axis. Draw the standard curve using coordinate paper or software, and determine the linear regression equation. Substitute the OD value of the sample into the equation to calculate its concentration.

Sample Preparation and Requirements

1. Serum: Allow whole blood collected in serum separation tubes to stand at room temperature for 2 hours or refrigerate overnight at 4°C. Centrifuge at 1000 × g for 20 minutes. Take the supernatant, and store it at -20°C or -80°C. Avoid repeated freezing and thawing.
2. Plasma: Collect specimens with EDTA or heparin as anticoagulant. Centrifuge within 30 minutes at 1000 × g for 15 minutes at 2–8°C. Store the supernatant at -20°C or -80°C, avoiding repeated freeze-thaw cycles.
3. Tissue Homogenization: Wash tissue with pre-cooled PBS (0.01M, pH=7.4) to remove residual blood. Weigh the tissue and add it to a glass homogenizer with PBS (typically 1:9 weight/volume ratio). Add protease inhibitors if needed. Grind on ice, sonicate, or freeze-thaw to lyse cells. Centrifuge at 5000 × g for 5–10 minutes and use the supernatant for testing.
4. Cell Culture Supernatant or Other Biological Samples: Centrifuge at 1000 × g for 20 minutes, take the supernatant, and store it at -20°C or -80°C. Avoid repeated freezing and thawing.

Note: Hemolyzed samples may affect the results. Do not test hemolyzed specimens.

Application

This kit is intended for the in vitro quantitative detection of superoxide dismutase (SOD) in serum, plasma, tissue homogenates, and other biological fluids.

Storage Conditions

Store the kit at 2–8°C. For optimal performance, keep the reagents sealed and protected from light.

Shelf Life

The shelf life of the kit is 6 months from the date of manufacture.