Below is a picture of ELISA results. Why is the color darker in some wells and lighter in others? Assume that all of these wells are test wells, not controls.
Below is a picture of ELISA results. Why is the color darker in some wells and lighter in others? Assume that all of these wells are test wells, not controls.
Basic Clinical Laboratory Techniques 6E
6th Edition
ISBN:9781133893943
Author:ESTRIDGE
Publisher:ESTRIDGE
Chapter4: Basic Immunology And Immunohematology
Section4.3: Tests For Rheumatoid Factors
Problem 2.1CS
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This image shows the results of an Enzyme-Linked Immunosorbent Assay (ELISA) conducted in a 96-well plate. The plate has 12 columns and 8 rows of wells. In this particular ELISA plate, you can observe wells that vary in color intensity from dark blue to very light blue or even clear.
### Explanation of Color Intensity
The color intensity in each well corresponds to the amount of target antigen or analyte present:
- **Darker Blue Wells:** These wells indicate a higher concentration of the target analyte. The enzyme linked to the antibody or antigen in the well has catalyzed a significant substrate conversion, leading to a strong color signal.
- **Lighter Blue or Clear Wells:** These wells indicate a lower concentration of the target analyte or the possible absence of it. Less substrate conversion has occurred, resulting in a weak color signal or no color development.
### Steps to Consider:
1. **Sample Addition:** Each well is initially coated with a sample containing the target antigen.
2. **Addition of Detection Antibody:** An antibody specific to the antigen, linked with an enzyme, is added.
3. **Substrate Addition:** A substrate for the enzyme is added, resulting in a color change.
4. **Reading the Plate:** The degree of color change is measured, usually using a spectrophotometer.
### Importance
Interpreting the color variations correctly is crucial in determining the presence and quantity of the target analyte in each sample. This information is vital for various diagnostic and research applications, such as detecting antibodies, hormones, or peptides in biological samples.
For a detailed analysis, the optical density (OD) readings from each well should be measured and compared to a standard curve to quantify the analyte concentration precisely.](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F2512f82d-69c0-4421-95ac-43df259f3b1a%2Fcd2704dc-1e96-4602-b677-ba2028fa3e1a%2Fdm71xne_processed.jpeg&w=3840&q=75)
Transcribed Image Text:**Understanding ELISA Results**
Below is a picture of ELISA results. Why is the color darker in some wells and lighter in others? Assume that all of these wells are test wells, not controls.
This image shows the results of an Enzyme-Linked Immunosorbent Assay (ELISA) conducted in a 96-well plate. The plate has 12 columns and 8 rows of wells. In this particular ELISA plate, you can observe wells that vary in color intensity from dark blue to very light blue or even clear.
### Explanation of Color Intensity
The color intensity in each well corresponds to the amount of target antigen or analyte present:
- **Darker Blue Wells:** These wells indicate a higher concentration of the target analyte. The enzyme linked to the antibody or antigen in the well has catalyzed a significant substrate conversion, leading to a strong color signal.
- **Lighter Blue or Clear Wells:** These wells indicate a lower concentration of the target analyte or the possible absence of it. Less substrate conversion has occurred, resulting in a weak color signal or no color development.
### Steps to Consider:
1. **Sample Addition:** Each well is initially coated with a sample containing the target antigen.
2. **Addition of Detection Antibody:** An antibody specific to the antigen, linked with an enzyme, is added.
3. **Substrate Addition:** A substrate for the enzyme is added, resulting in a color change.
4. **Reading the Plate:** The degree of color change is measured, usually using a spectrophotometer.
### Importance
Interpreting the color variations correctly is crucial in determining the presence and quantity of the target analyte in each sample. This information is vital for various diagnostic and research applications, such as detecting antibodies, hormones, or peptides in biological samples.
For a detailed analysis, the optical density (OD) readings from each well should be measured and compared to a standard curve to quantify the analyte concentration precisely.
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