1. ELISA Equipment

2. 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.

3. 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.

4. Substrate Primary antibody Enzyme Secondary antibody Different antigens in sample Coloured product

5. ELISA Qualitative/Quantitative Qualitative – determines antigen or antibody is present or absent Quantitative – determines the quantity of the antibody Titer

6. 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

7. Basic Steps Of E nzyme- L inked I mmuno s orbant A ssay

8. 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.

9. Using the equipment When 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

10. Equipment used in ELISA test

11. Maintenance and Calibration Schedule

13. PRINCIPLE OF INSTRUMENT

14. ELISA READER THERMOLAB SYSTEM (USA)