Tollip ELISA kits are used to measure the levels of Toll-interacting protein (Tollip) in biological samples. Tollip is involved in the regulation of inflammatory responses and immune system functions. Accurate measurement of Tollip levels can aid in research on immune system diseases and inflammatory conditions.

Components

• Coated Microplate: A 96-well plate pre-coated with antibodies specific to Tollip, allowing for the capture of Tollip from the sample.
• Standards: Recombinant Tollip proteins with known concentrations to create a standard curve for quantification.
• Detection Antibody: A secondary antibody specific to Tollip, usually conjugated to an enzyme like horseradish peroxidase (HRP) for detection.
• Substrate Solution: Contains a substrate that reacts with HRP, producing a color change proportional to the amount of Tollip.
• Stop Solution: An acid solution that halts the enzyme reaction and stabilizes the color for measurement.
• Wash Buffer: Used to wash away unbound proteins and reduce background interference.

Procedure

• Sample Preparation: Dilute and add samples and standards to the wells of the microplate.
• Incubation: Allow Tollip in the samples to bind to the antibodies on the plate.
• Detection: Add the detection antibody, which binds to the captured Tollip.
• Substrate Addition: Add substrate solution to develop a color proportional to Tollip levels.
• Stopping Reaction: Add stop solution to stop the color development.
• Measurement: Use a microplate reader to measure optical density (OD) and determine Tollip concentration based on the standard curve.

Technical Considerations

• Sensitivity: The minimum detectable concentration of Tollip.
• Specificity: Ability to specifically detect Tollip without interference from other proteins.
• Reproducibility: Consistency of results across different assays and conditions.
• Interferences: Potential impact of other substances in the sample on the accuracy of the assay.

Applications

• Research: Investigating Tollip’s role in immune regulation, inflammation, and related diseases.
• Clinical Diagnostics: Assessing Tollip levels in conditions linked to immune system dysregulation and inflammatory diseases.