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Summary: Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays, by using antibodies or antigens coupled to an easily-assayed enzyme. ELISAs can provide a useful measurement of antigen or antibody concentration. There are two main variations on this method: The ELISA can be used to detect the presence of antigens that are recognized by an antibody or it can be used to test for antibodies that recognize an antigen. An ELISA is a five-step procedure: 1) coat the microtiter plate wells with antigen; 2) block all unbound sites to prevent false positive results; 3) add antibody to the wells; 4) add anti-mouse IgG conjugated to an enzyme; 5) reaction of a substrate with the enzyme to produce a colored product, thus indicating a positive reaction. There are many different types of ELISAs. One of the most common types of ELISA is "sandwich ELISA."
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Protocols
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Affinity Measurements by Competition ELISA
(Dario Neri's labs, Institute of Pharmaceutical Sciences ETHZ)
The ELISA protocols for detection of the antibody binding to an antigen-coated microtitre plate are standard laboratory techniques. Most recombinant antibody fragments are typically detected using monoclonal antibodies directed against a peptidic tag engineered at the C-terminal extremity of the recombinant antibody. In certain experimental conditions, such peptidic tags may undergo proteolytic cleavage, thereby lowering the sensitivity of antibody detection. Reagents that bind to the antibody molecule without impairing antigen binding (e.g., protein A or protein L) may therefore be preferable. Alternatively, the experimental scheme described here can be performed in a similar fashion, using radiolabeled antibodies and radioacitve detection of antibody-mediated antigen binding.
https://www.pharma.ethz.ch/institute_groups/biomacr...
Added: Mon Sep 09 2002, Hits: 5410, Reviews: 0 Write review Cached -
Basic ELISA Protocol
(Immunology Applications Core Facility, University of Chicago)
Basic protocol for "sandwich ELISA"
https://iacf.bsd.uchicago.edu/MabHome/ELISA.htm
Added: Mon Apr 19 2004, Hits: 1598, Reviews: 0 Write review Cached - Cellular ELISA Protocol (Contributed by Nanci Donacki)
Added: Tue May 14 2002, Reviews: 0 Write review -
Competitive ELISA
(Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
https://www.nottingham.ac.uk/~mbzspd/methods/Compet...
Added: Sat Apr 10 2004, Hits: 688, Reviews: 0 Write review Cached -
ELISA (Enzyme-Linked ImmunoadSorbent Assay)
(Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
https://www.nottingham.ac.uk/~mbzspd/methods/ELISA....
Added: Sat Apr 10 2004, Hits: 1157, Reviews: 0 Write review Cached -
ELISA Inhibition Assay
(Goldberg Lab)
https://wheat.pw.usda.gov/~lazo/methods/goldberg/el...
Added: Tue May 14 2002, Hits: 1815, Reviews: 0 Write review Cached -
ELISA Procedure for Measuring Serum Antibody Titer
(Komabiotech)
the antigen (peptide or protein) is bound to the polystyrene microtiter plate first. The antiserum containing the anti-peptide antibody is then added to the well and allowed to bind. Finally, a second antibody, specific for the first antibody and labeled for detection, is added to the well and allowed to bind. The second antibody usually has an enzyme conjugated to it. This enzyme catalyzes the formation of colored substance from a colorless substrate. This colored substance is then quantified and the amount of antibody present can be calculated.
https://www.komabiotech.com/technical/protocol/ELIS...
Added: Tue May 14 2002, Hits: 4013, Reviews: 0 Write review Cached -
ELISA with Platelet
(Viral Immunity & Pathogenesis Group, the Université Catholique de Louvain)
This modification of qualitative ELISA (Enzyme-Linked Immunosorbent Assay) is used for either screening detection of anti-platelet antibodies or for detection of platelet-associated Ig (PAIg)
https://www.icp.ucl.ac.be/mexp/vipg_plelisa.html
Added: Wed Mar 24 2004, Hits: 2666, Reviews: 0 Write review Cached -
Experiment Data Sheet
(Contributed by Nanci Donacki)
Useful sheet for recording ELISA data
http://www.protocol-online.org/protocols/experimen...
Added: Tue May 14 2002, Hits: 3927, Reviews: 0 Write review - Gangliosides ELISA protocol (Contributed by Pingsunjim)
This protocol can be used for detection of gangliosides.Spicific antibodies to gangliosides are added to cells and allowed to bind to the cell surface. The nonbound antibody is washed away and an enzyme conjugated second antibody is then detected by the addition of an enzyme-spicific substrate.
Added: Tue May 14 2002, Reviews: 0 Write review - Live Cell ELISA Protocol ( Contributed by Nanci Donacki )
Added: Tue May 14 2002, Reviews: 0 Write review - Peptide Inhibition ELISA Protocol (Contributed by Nanci Donacki)
Added: Tue May 14 2002, Reviews: 0 Write review - Periodate Treatment of Fixed Cell Plates (Contributed by Nanci Donacki)
Added: Tue May 14 2002, Reviews: 0 Write review
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