Laboratory #11 - ELISA

A series of flashcards created for reviewing the ELISA laboratory procedure and concepts related to the study of the Foxvirus.

What is the objective of Laboratory #11 - ELISA?

To successfully identify 'Patient Zero' by utilizing ELISA to determine infected individuals and the distribution of the disease.

What virus is the focus of this ELISA lab?

The Foxvirus.

What type of ELISA is being performed in this lab?

A direct ELISA.

What happens to the antigen if it is present in the sample during a direct ELISA?

It will bind to the plate.

What type of enzyme is used in the antibody for the ELISA?

Horseradish peroxidase (HRP).

What is the substrate used in the ELISA procedure?

TMB (3,3',5,5'-Tetramethylbenzidine).

What indicates that the Foxvirus antigen is present in the sample?

A color change occurs when the substrate is catalyzed by the enzyme.

What is a positive control sample used for in the lab?

It confirms that the ELISA test is functioning correctly by changing color.

What should happen to the negative control sample during the ELISA test?

It should not change color.

What materials are needed for the ELISA procedure?

Samples, positive control, negative control, antibodies, and substrate.

How many exchanges should be made during Exercise 11, Part 1?

A total of 4 exchanges.

What does the procedure simulate when students exchange droppers?

The spread of infection through handshakes, sneezing, or coughing.

What must students record during the exchanges in Exercise 11, Part 1?

The names of the students with whom they exchanged fluids.

What will indicate that the antibody has bound to the Foxvirus in the ELISA?

A color change in the sample as a result of substrate catalysis.

What does ELISA stand for?

Enzyme-Linked Immunosorbent Assay.

What is the role of antibodies in the ELISA procedure?

To detect the presence of the specific antigen by binding to it.

What should a student do if the antigen is not present in their sample?

There will be no binding and no color change will occur.

What is the significance of using HRP labeled primary antibodies?

They enable the detection of the antigen through a color change when substrate is catalyzed.

What is the expected result for the 'positive' control in the ELISA?

It should exhibit a color change.

What is the expected result for the 'negative' control?

It should not exhibit any color change.

How many drops of the Foxvirus antibody are added to each tube?

5 drops.

How many drops of substrate are added to each tube in the ELISA?

10 drops.

During the ELISA procedure, what does color change signify?

The presence of the Foxvirus antigen.

What characteristic of the Foxvirus antibody allows it to be detected?

It has a color-changing chromatophore attached.

What action must be avoided when exchanging fluid with other students?

Do not exchange fluid with the same person more than once.

What will students observe in Exercise 11?

The differences in color change between their samples and control samples.

What is the primary goal of performing the ELISA in this lab?

To determine who in the class has been infected and to identify 'Patient Zero'.

Why is it important to compare samples to positive and negative controls?

To validate the results and the function of the ELISA assay.

What safety protocol should be followed during the lab?

Proper handling of samples to prevent contamination.

What does a lack of color change in a patient sample indicate?

The absence of the Foxvirus in that sample.

In the ELISA procedure, why is a conjugated enzyme necessary?

To enable the detection of antigen-antibody complexes through a measurable color change.

What type of study does this ELISA experiment simulate?

An epidemiological study to track the spread of infection.

What is the functionality of horseradish peroxidase in the ELISA test?

It catalyzes the reaction with the substrate to produce a color change.