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

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Kaixin micro test

Test probe P100-M3

Experimental Principle

This 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 sequentially, the plate is incubated and thoroughly washed. A TMB substrate is then added, which turns blue under peroxidase catalysis and finally changes to yellow when an acid is introduced. The intensity of the color is directly proportional to the SOD concentration in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader, allowing for accurate calculation of the sample concentration.

Kit Composition

Result Calculation

To calculate the results, plot the OD values of the standards against their known concentrations on graph paper or using software. Obtain the linear regression equation from the standard curve, and use it to determine the concentration of the unknown samples by substituting their OD values into the equation.

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 at -20°C or -80°C. Avoid repeated freezing and thawing.
2. Plasma: Collect specimens using EDTA or heparin as anticoagulant. Centrifuge at 1000 × g for 15 minutes within 30 minutes of collection at 2–8°C. Store the supernatant at -20°C or -80°C, avoiding repeated freeze-thaw cycles.
3. Tissue Homogenate: Wash tissue with pre-cooled PBS (0.01M, pH=7.4) to remove blood. Weigh the tissue, add an appropriate volume of PBS (e.g., 1g:9mL), and homogenize on ice. Add protease inhibitors if needed. 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, collect the supernatant, and store at -20°C or -80°C. Avoid repeated freezing and thawing.

Note: Hemolyzed samples may interfere with the results and should not be used.

Application

This kit is designed for the quantitative in vitro detection of superoxide dismutase (SOD) in serum, plasma, tissue homogenates, and other related liquid samples.

Storage Conditions

Store the kit at 2–8°C. For optimal performance, ensure the reagents are kept in a stable environment away from moisture and light.

Shelf Life

The shelf life of the kit is 6 months when stored properly.