ELISA Equipment

INTRODUCTION TO ELISA

• ELISA, or Enzyme-Linked Immunosorbent Assay, are immunological procedures used to determine the presence of antibodies or specific antigens in samples.

• The technique is based on the detection of an antigen or antibodies captured on a solid surface using direct or secondary, labeled antibodies, producing a colored reaction whose product can be read by the spectrophotometer.

Basic principle of ELISA

• Use an enzyme to detect the binding of antigen (Ag) antibody (Ab). The enzyme converts a colorless substrate to a colored product, indicating the presence of Ag : Ab binding.

• ELISA can be used to detect either the presence of Antigens or antibodies in a sample depending how the test is designed.

Substrate

Primary antibody

Enzyme

Secondary antibody

Different antigens in sample

Coloured product

ELISA Qualitative/Quantitative

• Qualitative– determines antigen or antibody is present or absent

• Quantitative – determines the quantity of the antibody Titer

ELISA Components• Coated Plates (The 96-well coated with either inactivated antigen or

an antibody.)• Controls ( Positive and negative)• Conjugate : Enzyme-labeled antibodies or antigens that react

specifically to plate-bound sample analytes• Substrate: reacts with the enzyme portion of the conjugate to • produce color.• Wash Concentrate: Used to wash away unbound materials from the

plates.• Stop solution: stops the enzyme-substrate reaction

Basic Steps Of Enzyme-Linked Immunosorbant

Assay

Equipment required for ELISA testing

• In order to perform the ELISA technique, the following equipment is required:

1. Micro plate reader. 2. Micro plate washer. 3. Liquid dispensing system (multi-channel

pipettes may be used). 4. Incubator to incubate the plates.

Using the equipmentWhen an ELISA test is conducted, it typically follows these steps:1. A first washing of the plate may be done using the microplate washer.

2. Using a liquid dispenser or the multi-channel pipettes, wells are filled with the solution prepared to be used in the test.

3. The plate is placed in the incubator where at a controlled temperature, a series of reactions take place.

4. Finally, when all the incubation steps have been completed, the plate is transferred to the microplate reader

Equipment used in ELISA test

Maintenance and Calibration Schedule

PRINCIPLE OF INSTRUMENT

ELISA READER

THERMOLAB SYSTEM (USA)