ELISA Methods, Techniques & Protocols




Definition: enzyme-linked immunosorbent assay (ELISA) is a technique for detecting the presence of specific substances, such as enzymes, viruses, bacteria, or antibodies in blood.

General Methods & Techniques

Antigen-Antibody Specific Applications

Tissue-Cell Specific Applications

Virological Applications

Tumor, Disease & Diagnostic Applications

Basic ELISA Protocol - There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common types of ELISA is the so-called "sandwich ELISA." It is termed this because the antibody that you are detecting gets sandwiched between an antigen and a chromogenically-conjugated antibody. Below you will find a basic protocol for this assay.

ELISA (Enzyme-Linked Immunosorbent Assay) Home Page - This page is dedicated for researchers having some troubles in making ELISA, especially for non-experts. If you have troubles with ELISA, go to check the trouble shootings and please ask me directly.

Detection of Recombinant Proteins By ELISA - Refer to the Western Blot Protocol for preparation of bacterially expressed protein. From a 10 ml bacterial culture, 2-4 plates can be coated for a direct ELISA.

Monoclonal Antibody Screening by Direct ELISA - This is only a semiquantitative assay. If you want a quantitative assay, you have to  dilute the primary antibody (step 4) at least 1/20 in blocking solution and probably much more. Anyway, direct ELISAs are adequate for quantitative assays only with pure samples. In any other case, you should use indirect ELISAs or sandwich ELISAs.

ELISA Inhibition Assay - Goldberg Lab Standard Protocols

ELISA Procedure for Measuring Serum Antibody Titer - Immunoassays are a powerful technique for detecting and measuring antigens and antibodies.  Immunoassays can be classified three ways based on the steps involved.

ELISA with Platelets Protocol - 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) (shown here).