Investigating Human Health Using the ELISA
This is a replay of our biotechnology training workshop which was held at 2:00PM ET on Thurs, Feb 24.
The Enzyme-Linked Immunosorbent Assay, or ELISA, is a highly sensitive test that uses antibodies to detect the presence of specific molecules within a complex sample. Since we can generate antibodies to lots of different molecules, the ELISA has been adapted for many uses. In this workshop, we will perform a simple ELISA that comes with several scenarios, including pregnancy testing, early detection of heart attacks, and identification of gluten in food products. This allows you to tailor the experiment to fit seamlessly within your curriculum!
For the experiment: https://www.edvotek.com/279_1
For the slides: www.edvotek.com/site/pptx/edvotek-youtube-human-health-elisa-279.pptx
______________________________
Don't forget to like and subscribe to our social media!
YouTube: / edvotekinc
Twitter: / edvotek
Instagram: / edvotek
/ edvotek
TikTok: / edvotek
_______________________________
The Enzyme-Linked Immunosorbent Assay, or ELISA, is a highly sensitive test that uses antibodies to detects the presence of specific molecules within a complex sample. The test we are running today is a real ELISA, in that it uses real antibodies to detect specific molecules in samples. But, we’re not using any samples that are infectious. Instead, we are using non-hazardous reagents that allow students to perform an authentic ELISA and applying different scenarios to the interpretation of the results. The lessons provided in this kit focus on common tests used in medicine and food production. This allows you to choose a scenario that fits best into your curriculum.
Since we can generate antibodies to lots of different molecules, the ELISA is highly versatile laboratory test that has been adapted for many uses. For example, you may have an allergy, so you need to be careful about what you eat. Foods that may contain common allergens can be tested by ELISA to indicate that they are safe to eat. For this workshop, we’ll be discussing the ELISA as it is used in medical testing. The ELISA is commonly used for medical diagnostics, as it is can be used to identify antigens in blood, saliva, urine and other biological samples. If you’ve ever taken a pregnancy test, you’ve already performed an ELISA at home.
The ELISA can be qualitative or quantitative. The qualitative ELISA gives us a Yes or No answer: A strong signal is positive, no signal is negative. One example of a qualitative ELISA that you may be familiar with is a pregnancy test, which we will discuss more later. This at-home ELISA tests urine for a hormone that is an indicator of pregnancy. You can’t be just a little bit pregnant! When taken at the right time, a pregnancy test will give you a yes or no answer.
In the Qualitative ELISA, we compare results from unknown to results from samples of known concentration. (Advance slide) So back to that example of food allergens or gluten. The ELISA is often used to test for the presence of known allergens in food that may be introduced through manufacturing. Here’s a label from some certified gluten-free oats. You can see from the label that they certify to 3ppm, or parts per million.
There are many protocols for the ELISA, but they all rely on using antibodies to detect the presence of antigens in experimental samples, and they follow the same basic principles. First, we add our sample, where it sticks to the plastic walls of our microtiter plate through hydrophobic and electrostatic interactions.
After washing away any excess sample, the wells are “blocked” with a protein-containing buffer to prevent non-specific interactions. This is important, because proteins are also antibodies that can stick to the wells. We then add the primary antibody to each well. If the antigen is present, the antibody binds through electrostatic interactions. Excess antibody is washed out of the wells. In this ELISA, our antibodies come from the patient samples
The secondary antibody, which recognizes the primary antibody, is added to the wells. If the antibody-antigen complex has formed in the well, it will remain in the well after the wells are washed.
The secondary antibody is covalently linked to an enzyme that allows for the detection of the antibody-antigen complex. A clear, colorless substrate solution is added to each well. In wells where the secondary antibody is present, the enzyme turns the clear substrate solution to blue. Since most enzymes have a high catalytic activity, meaning that they can quickly turn over the substrate, this assay allows us to quickly detect even the smallest amount of antibody.
Смотрите видео Investigating Human Health Using the ELISA онлайн без регистрации, длительностью часов минут секунд в хорошем качестве. Это видео добавил пользователь Edvotek Inc. 01 Январь 1970, не забудьте поделиться им ссылкой с друзьями и знакомыми, на нашем сайте его посмотрели 1,25 раз и оно понравилось 1 людям.