This document provides an overview of the ELISA (Enzyme-Linked Immunosorbent Assay) procedure. It discusses the basic principle, history, equipment needed, types (direct and indirect), precautions, advantages/disadvantages, limitations, and applications of ELISA. The procedure involves coating a 96 well plate with an antibody, adding samples and secondary antibodies conjugated with reporter molecules, washing unbound molecules, and adding a substrate to develop color if the target antigen is present in the sample.

2. Presenters
Aneesa Maaz 13
Mahnoor Anwar 54
Kainat Ayub 60

4. Contents
• INTRODUCTION
• DEFINITION
• BASIC PRICIPLE OF ELISA
• HISTORY
• EQUIPMENTS
• PROCEDURE
• TYPES(direct, indirect)
• PRECAUTIONS
• ADVANTAGES AND DISADVANTAGES
• LIMITATIONS
• APPLICATIONS

17. procedure
*Coat 96 well plate with suitable antibody or
antisera....(2 hours or over night)
*wash the wells for three times 5,5 mints with
PBS..(phosphate buffer salline)
*bloock the wells with 10% BSA or skim milk
protien...(1 hr room temp)
*washing again
*Add sample of your taste....(1 hour ,room
temp)

18. • *the antibody you coated to plate will have
complimentarity to sampl so it wil get bind.
• *washing
• *secondry antibody is added conjugated with
reporter molecule with the help of
biotin...(enzyme or radioactiv molecule)
• Biotin::::>stiptividin::::::>HRPO:::::::;
• *wash to remov unbind molecule

19. • *now add substrate...(TMB) to the solution
which develop a colour
• *if the test sample contain the antigen the
colour will develop if not present colour will
not develop