Tag: immunology
ELISA: Indirect, Blocking, Capture Format and Components
Sensitive and reproducible technologies available. These assays are rapid, simple to perform, and easily automated. IDEXX introduced the first commercial poultry ELISA for infectious bursal disease (IBD) in 1985 and the first commercial livestock ELISA for Aujeszky’s disease/pseudorabies in 1986, enhancing the way laboratories test production animals. As with any assay, the reproducibility and reliability…
ELISA Equipment, Maintenance and Calibration
Equipment for ELISA testing is widely available. Readers, washers, and pipettes are available as manual or automated systems. Some of the factors affecting equipment selection are the number and types of tests and samples, technical training of staff, and financial considerations. Below is a brief outline of some equipment available for performing ELISA testing. Pipettes…
ELISA FAQs
ELISA FAQs The ELISA protocols do not recommend shaking during incubations. Have you tested shaking and decided against it or is it unnecessary? We tested our protocols with and without shaking during incubations and determined that there is no difference between the two approaches. Therefore, we believe that shaking is not necessary. Is your ELISA…
Indirect ELISA – Antigen Coating, Blocking, Preparation
Indirect ELISA – This is a general protocol in which antigen coating and blocking may not be required if the wells from the manufacturer have been pre-adsorbed with the antigen. 1. Antigen Coating Dilute purified antigens to a final concentration of 1-10 μg/mL in bicarbonate/carbonate antigen-coating buffer (100 mM NaHCO3 in deionized water; pH adjusted…
Sandwich ELISA – Antibody Coating, Blocking, Preparation
Sandwich ELISA – All of the ELISA kits from Boster use the sandwich format and avidin-biotin chemistry. Our ELISA assays require the dilutions of standard solutions, biotinylated antibody (detection antibody) and avidin-biotinperoxidase. 1. Capture Antibody Coating (These steps are not required if the pre-adsorbed Picokine ELISA kits from Boster are used) Dilute the capture antibody…
Competitive ELISA : Antigen Coating, Blocking, Preparation
Competitive ELISA, This is a general protocol in which antigen coating and blocking may not be required if the wells from the manufacturer have been pre-adsorbed with the antigen. 1. Antigen Coating Dilute purified antigens to a final concentration of 20 μg/ml in bicarbonate/carbonate antigen-coating buffer (100 mM NaHCO3 in deionized water; pH adjusted to…
ELISA Test (enzyme-linked immunosorbent assay)
ELISA Test – ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying peptides, proteins, antibodies, and hormones. In an ELISA, an antigen must be immobilized to a solid surface and then complexed with an antibody that is linked to an enzyme. Detection is accomplished by assessing the conjugated enzyme activity…