What is an ELISA Kit and What are the Popular ELISAs?



ELISAs are kinds of immunoassays highly practiced to estimate the quantity of a target within the sample. Thus, more and more researchers are looking for ELISA kit manufacturers in India. The samples that can be tested in a good ELISA include serum, plasma, cell culture supernates, cell lysates, saliva, tissue lysates, and urine. There are many immunoassay platforms for measuring and quantifying protein levels in biological fluids. In most cases, ELISAs remain preferred for increased sensitivity and specificity, and higher accuracy tolerance of harsh buffers or pretreatments. This article will discuss everything that you need to know about ELISA.

What is an ELISA Kit?

An ELISA kits distributor will supply the best ELISA. These ELISAs are carried out in 96-well microplates that are precoated with a capture antibody specific to the target analyte of interest. After incubation with experimental samples, standards or controls, this capture antibody captures the target analyte. The completion of the sandwich is by a conjugated detection antibody to another epitope on the target analyte. A subsequently added substrate solution then produces a signal directly proportional to the amount of analyte bound. ELISAs exist in different formats.

Types of ELISAs

The popular types of ELISAs are as follows:

Direct ELISA

In a direct ELISA bought from reputed ELISA kit suppliers, the antigen or sample is immobilized directly on the plate, and the target protein is detected with a conjugated detection antibody. Then, the substrate is added to produce a signal that is proportional to the amount of analyte in the sample. In a direct ELISA, because only one antibody is used, it is less specific than a sandwich ELISA. It is used to assess antibody affinity and specificity. Also, it helps investigate blocking/inhibitory interactions.

Indirect ELISA

An indirect ELISA is a direct ELISA, except the antigen immobilized on a plate has an additional amplification detection step. In indirect, first, an unconjugated primary-detection antibody is added and binds to a specific antigen, and then the conjugated secondary antibody against the host species of primary antibody is added. The substrate then produces a signal proportional to the amount of antigen bound in the well. It helps in the measurement of endogenous antibodies.

Sandwich ELISA

The most widely used ELISA is the sandwich ELISA. The type of antibody pairs used is often referred to as matched antibody pairs and sandwiches the antigen. A microplate coated with the capture antibody has the sample added to it and subsequently binds and is immobilized on the plate by the protein of interest. In the next step, a conjugated-detection antibody is added, which binds to another epitope on the target protein. The substrate is added and produces a signal proportional to the amount of analyte present in the sample. Since two antibodies have to associate with the binding of the protein of interest, sandwich ELISAs are very specific. It helps to find out the concentration of analyte in a biological sample.

Competitive ELISA

It is one of the most commonly used ELISAs for small molecules in the case where the protein of interest is simply too small to efficiently sandwich with two antibodies. The ELISA kit suppliers in India supply this kit the most. This is another version of the sandwich ELISA but instead of having the detection antibody conjugated, it uses a conjugated antigen to compete for binding with an antigen present in the sample. The more antigens in the sample, the fewer conjugated antigens can bind with the capture antibody. Then substrate is added and the signal produced will be inversely proportional to the amount of protein content in the sample. It helps in determining the concentrations of small molecules and hormones.

Conclusion

ELISA, the acronym for Enzyme-linked Immunosorbent Assay, is a laboratory test that detects and measures the quantity of a substance in a sample. ELISAs are applied to determine and quantify antibodies, antigens, proteins, and hormones. If you want to know the ELISA kits price, just reach out to Universal Biotechnology as they offer high-quality kits at affordable prices.

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