Scribd
Upload
57 views1 page

ELISA

ELISA is a biochemical technique used to detect antibodies or antigens in a sample. It works by affixing an unknown amount of antigen to a surface, then applying a specific antibody linked to an enzyme. A detectable signal is produced when a substrate is added that the enzyme can convert. The amount of antigen in the sample can be inferred from the magnitude of the signal, such as fluorescence. Performing ELISA involves immobilizing the sample on a plate and forming complexes between antigens and detection antibodies, which may be linked directly to an enzyme or detected by a secondary antibody linked to an enzyme. Washing and developing the plate produces a visible signal indicating the quantity of antigen in the sample.

Uploaded by

phvega06
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
57 views1 page

ELISA

ELISA is a biochemical technique used to detect antibodies or antigens in a sample. It works by affixing an unknown amount of antigen to a surface, then applying a specific antibody linked to an enzyme. A detectable signal is produced when a substrate is added that the enzyme can convert. The amount of antigen in the sample can be inferred from the magnitude of the signal, such as fluorescence. Performing ELISA involves immobilizing the sample on a plate and forming complexes between antigens and detection antibodies, which may be linked directly to an enzyme or detected by a secondary antibody linked to an enzyme. Washing and developing the plate produces a visible signal indicating the quantity of antigen in the sample.

Uploaded by

phvega06
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

ELISA

 Enzyme-linked immunosorbent assay (ELISA), also known as an enzyme


immunoassay (EIA), is a biochemical technique used mainly in immunology to detect
the presence of an antibody or an antigen in a sample. The ELISA has been used as a
diagnostic tool in medicine and plant pathology, as well as a quality control check in
various industries. In simple terms, in ELISA, an unknown amount of antigen is affixed to
a surface, and then a specific antibody is applied over the surface so that it can bind to
the antigen. This antibody is linked to an enzyme, and in the final step a substance is
added that the enzyme can convert to some detectable signal. Thus in the case of
fluorescence ELISA, when light of the appropriate wavelength is shone upon the sample,
any antigen/antibody complexes will fluoresce so that the amount of antigen in the
sample can be inferred through the magnitude of the fluorescence.

 Performing an ELISA involves at least one antibody with specificity for a particular
antigen. The sample with an unknown amount of antigen is immobilized on a solid
support (usually a polystyrene microtiter plate) either non-specifically (via adsorption to
the surface) or specifically (via capture by another antibody specific to the same antigen,
in a "sandwich" ELISA). After the antigen is immobilized the detection antibody is added,
forming a complex with the antigen. The detection antibody can be covalently linked to
an enzyme, or can itself be detected by a secondary antibody which is linked to an
enzyme through bioconjugation. Between each step the plate is typically washed with a
mild detergent solution to remove any proteins or antibodies that are not specifically
bound. After the final wash step the plate is developed by adding an enzymatic substrate
to produce a visible signal, which indicates the quantity of antigen in the sample.

 Traditional ELISA typically involves chromogenic reporters and substrates which


produce some kind of observable color change to indicate the presence of antigen or
analyte. Newer ELISA-like techniques utilize fluorogenic, electrochemiluminescent, and
real-time PCR reporters to create quantifiable signals. These new reporters can have
various advantages including higher sensitivities and multiplexing. Technically, newer
assays of this type are not strictly ELISAs as they are not "enzyme-linked" but are
instead linked to some non-enzymatic reporter. However, given that the general
principles in these assays are largely similar, they are often grouped in the same
category as ELISAs.

You might also like