ELISA Kits Principle

Basic Principles of ELISA Kits

The basis of ELISA is the solid phase of antigen or antibody and enzyme labeling of antigen or antibody. Although the antigen or antibody bound to the surface of the solid phase carrier still maintains its immunological activity, the enzyme-labeled antigen or antibody retains both immunological activity and enzyme activity.

In detection, the substance under test (antigen or antibody) in the sample binds to immobilized antigens or antibodies. Wash the plate to remove non-conjugates, and then add enzyme-labeled antigens or antibodies. At this time, the amount of enzyme that can be immobilized is related to the amount of the substance to be tested in the sample. After adding the substrate that reacts with the enzyme to develop color, the content of the substance in the sample can be judged according to the depth of the color, and qualitative or quantitative analysis can be carried out. Due to the high catalytic efficiency of the enzyme, the result of the immune reaction is indirectly amplified, so that the determination method can achieve high sensitivity.

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Components of ELISA Kits

• ELISA plates
  ELISA plates serve as solid support for immobilizing the captured antibodies and detecting the analytes of interest. These plates are typically flat-bottomed and come in various formats, such as 96-well, 384-well, and 1536-well plates. The choice of plate format depends on the specific requirements of the assay, including the number of samples and the level of throughput. We provide high-quality ELISA plates made from polystyrene or polyvinyl chloride, ensuring optimal performance and reliability.
• Standards and controls
  ELISA kits include a set of standards with known concentrations of the analyte, allowing researchers to generate a standard curve for quantitative analysis. These standards help determine the concentration of the analyte in the samples. Additionally, controls are included in the kit to validate the assay performance and ensure accurate results.
• Capture and detection antibodies
  ELISA kits consist of both capture and detection antibodies. The capture antibodies are immobilized onto the surface of the ELISA plate, where they specifically bind to the target analyte. The detection antibodies are labeled with enzymes or fluorophores and are used to detect the bound analyte. The choice of antibodies is critical for the sensitivity and specificity of the assay. We offer a wide selection of high-quality antibodies validated for their performance in ELISA assays.
• Conjugates and substrates
  To visualize and quantify the analyte-antibody complexes, ELISA kits include conjugates and substrates. Conjugates are typically enzyme-labeled secondary antibodies that bind to the detection antibodies. These conjugates enable the enzymatic or fluorescent detection of the analyte. Substrates are specific to the conjugated enzyme and produce a measurable signal, such as a color change or fluorescence, upon enzymatic reaction. The choice of conjugates and substrates depends on the detection system employed in the assay.
• Wash buffers and diluents
  ELISA kits also provide specialized wash buffers and diluents. Wash buffers are used to remove unbound substances and minimize background noise. They ensure the specificity and sensitivity of the assay by reducing non-specific binding. Diluents, on the other hand, are used to prepare the standards, controls, and samples at appropriate concentrations for analysis. These buffers and diluents are optimized to enhance the performance of the assay and improve the signal-to-noise ratio.
• Detailed protocol and technical support
  Our ELISA kits come with detailed protocols, providing step-by-step instructions on sample preparation, reagent handling, and assay procedures. In addition, our dedicated technical support team is available to answer any questions and provide expert guidance throughout the experimental process, ensuring researchers achieve reliable and reproducible results.