ELISA Disease Testing

Part 1: Introduction

This lab will let us test if we have a contagious "disease." The ELISA method tests for the presence of antibodies specific to a disease. So if you have the disease, the antibodies will be present, and if you don't, then the antibodies will be absent. This method has had a huge impact on the medical world. It is currently commonly used for disease detection, pregnancy tests, detecting drug use, testing air quality, and determining if food is actually what it's labeled.

Part 2: Experiment

First, we will label a yellow tube and pipet with our initials. Then we'll transfer our bodily fluid sample into another student's tube, then mix the samples, and take back half (750 microliters) and put it in our own tube, and write down their name and the time it was collected (including seconds.) Then we'll repeat the sharing protocol two more times when instructed, and discard the pipet. Then we'll store the tubes overnight.
On the next day, we'll label our 12-well strip as indicated in the manual. Then we'll use a fresh pipet tip to transfer 50 microliters of the positive control into the 3 positive wells, and use another fresh pipet tip to put 50 microliters of the negative control into the 3 negative wells. Then we'll wait 5 minutes, then wash the microplate strip on the paper towels. Then we'll use a new transfer pipet to fill each well with wash buffer, then drain it again on the paper towels. Then we'll do this washing step again. After that we'll transfer 50 microliters of primary antibody into all 12 wells, then wait 5 minutes, and wash twice again. Next we'll use a new pipet tip to transfer 50 microliters of secondary antibody into all 12 wells, awit 5 minutes, and wash 3 times. Then using a new pipet tip, we'll put 50 microliters of enzyme substrate into all 12 wells. Then we'll wait 5 minutes, and observe and record the results. Then we clean up, and we're done!