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Indirect ELISA Kit
The Indirect ELISA Kit is a widely used assay system designed for the detection and quantification of specific analytes such as proteins, antibodies, and other biomolecules within complex biological samples. This kit utilizes a two-step detection process that enhances sensitivity and flexibility compared to the direct ELISA method.
 
In an indirect ELISA, the process begins with the immobilization of the target antigen on the surface of the plate. A primary antibody, specific to the antigen, is then added to bind the target. After a washing step to remove unbound antibodies, a labeled secondary antibody is introduced. This secondary antibody is designed to recognize and bind to the host species of the primary antibody. The secondary antibody is typically conjugated to an enzyme, such as horseradish peroxidase (HRP) or alkaline phosphatase (AP). Following a second wash step to remove unbound secondary antibodies, a substrate is added. The enzyme acts on the substrate to produce a color change, which can be measured to determine the presence and quantity of the target analyte.
 
Advantages of Indirect ELISA:

  • Higher Sensitivity: The use of a secondary antibody amplifies the signal, making indirect ELISA more sensitive than direct ELISA, allowing for the detection of lower concentrations of the target analyte.
  • Cost-Effective: Since the secondary antibodies are often commercially available and can be used with a variety of primary antibodies, this method is generally less expensive than direct ELISA.
  • Flexibility: The method is versatile, as it allows the use of many different primary antibodies, making it suitable for a wide range of applications.
 
Disadvantages of Indirect ELISA:

  • Risk of Cross-Reactivity: The primary disadvantage is the potential for cross-reactivity between the secondary antibody and other proteins in the sample, which can lead to nonspecific signals and reduced accuracy.
 
The Indirect ELISA Kit is particularly useful for applications where high sensitivity is required and flexibility in antibody selection is advantageous.


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Sample: Serum Species:
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