Final Exam Flashcards
What is an ELISA, and what does ELISA stand for? Why is it used? What are some common applications of an
ELISA test?
ENZYME LINKED IMMUNOSORBENT ASSAY
It is used to detect and measure the presence of specific substances such as antibodies and antigens in a sample.
Common applications - pregnancy test or hiv test
How are enzymes important to ELISA’s?
They help detect and measure specific molecules accurately in a sample.
What is the purpose of the substrate in an ELISA test?
To react with the enzyme and produce detectable enzymes.
Why is there a primary and a secondary antibody? What is the purpose of each in an ELISA?
The primary antibody is the specific antibody that recognizes the antigen.
The secondary antibody recognizes the primary antibody.
They work together to increase the sensivity of the ELISA test
What is an antigen? What is an antibody? How do they relate to each other and to an ELISA
Antigen= specific foreign molecules that trigger an immune response in our bodies.
Antibody= a type of protein to help neutralize harmful susbatances such as bacteria and viruses.
They relate to each other and an ELISA because when antigens enter our bodies it stimulates the production of specific antibodies that can recognize and bind to antigens.
The binding of antibodies to antigens allows for the detection of the target molecule in the ELISA assay.
Why did we run the assay tests in triplicate?
To get a more accurate result
To account for potential errors for each try
What is the purpose of using positive and negative controls in an ELISA test?
To help confirm the test accuracy
Positive control contains what substance is being detected
Negative control does not contain the substance and should always be negative.
How does an ELISA work (what are the different components involved)?
Step 1= antigen
Step 2= Primary antibody
Step 3= Enzyme labeled secondary antibody
Step 4= chromogenic enzyme
It uses components of the immune systems and chemicals for detection.
What are the different levels of protein structure? How does this relate to an enzyme and its ability to function?
Protein structure:
Primary- sequence of amino acids in a protein
Secondary- folding of the polypeptide chain into regular patterns like beta sheets
Tertiary- overall three dimensional shape of a single polypeptide chain
Quaternary- the arrangement of multiple polypeptide chains to form a functional protein complex
The maintenance of the proteins structures are essential for an enzyme to function properly because it would affect the enzymes active site altering its ability to bind to substrates.
Understand the relationship between enzymes, substrates, and products
Enzymes act as catalyst to speed up the conversion of substrates into products.
What was the purpose of each of the reagents used in the experiment? (Properties of enzymes)
H202- used to clean and disinfect
Peroxidase— quickly eliminates h202 to reduce damage
Guaiacol- used for an indicator of oxygen
Ph5 buffer- maintains ph
What are some environmental factors that can affect the structure of a protein (enzyme)?
Temperature( if its to high) it disrupts the bonds that maintain the structure
pH levels (alter the change on amino acids)
What does it mean if an enzyme is denatured? If the rate is low, does it always mean that the enzyme was denatured?
Why or why not?
Denatured means- the structure has been disrupted and loses its normal shape and functions. Due to factors such as high temp and ph levels.
NO, there can be other factors and desaturation is just one possible cause of reduced enzyme activity.
Why is peroxidase important to living organisms?
Eliminates h20 TO REDUCE DAMAGE
It acts as a signaling molecule to regulate cellular processes like cell growth, immune responses and wound healing.
- How was the rate of the enzyme calculated in these experiments?(PROPERTIES OF ENZYMES)
