Overview: 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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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.
http://www.pharma.ethz.ch/institute_groups/biomacr...
Added: Mon Sep 09 2002, Hits: 4064, Reviews: 0 Write review Cached -
Basic ELISA Protocol
(Immunology Applications Core Facility, University of Chicago)
Basic protocol for "sandwich ELISA"
http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_F...
Added: Mon Apr 19 2004, Hits: 18126, 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)
http://www.nottingham.ac.uk/~mbzspd/methods/Compet...
Added: Sat Apr 10 2004, Hits: 5713, Reviews: 0 Write review Cached -
Detection of Recombinant Protein by ELISA
(Lab of Dr. Mark Barton Frank, Oklahoma Medical Research Foundation)
Direct ELISA method of detection of recombinant protein in bacteria
http://omrf.ouhsc.edu/~frank/ELISA.html
Added: Thu Sep 01 2005, Hits: 4059, Reviews: 0 Write review Cached -
Direct ELISA Protocol Using Fluorescent Substrate
(Abcam)
http://www.abcam.com/ps/pdf/protocols/Direct%20ELI...
Added: Wed Jan 18 2006, Hits: 2830, Reviews: 0 Write review -
Direct ELISA Protocol Using Primary Antibody
(Abcam)
http://www.abcam.com/ps/pdf/protocols/Direct%20ELI...
Added: Wed Jan 18 2006, Hits: 4326, Reviews: 0 Write review -
ELISA
(The Minion Lab, College of Veterinary Medicine at Iowa State University)
ELISA and conjugate preparation procedure.
http://mycoplasmas.vm.iastate.edu/lab_site/methods...
Added: Tue Nov 16 2004, Hits: 8261, Reviews: 0 Write review Cached -
ELISA (Enzyme-Linked ImmunoadSorbent Assay)
(Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
http://www.nottingham.ac.uk/~mbzspd/methods/ELISA....
Added: Sat Apr 10 2004, Hits: 8920, Reviews: 0 Write review Cached -
ELISA Inhibition Assay
(Goldberg Lab)
http://wheat.pw.usda.gov/~lazo/methods/goldberg/el...
Added: Tue May 14 2002, Hits: 4654, Reviews: 0 Write review Cached