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)
- What happened to the rate of peroxidase at different temperatures? Why?
As the temp increases the rate of peroxides temperatures increases as well
- What happened to the rate of peroxidase when other conditions were varied? Why?
What is the overall equation for cellular respiration?
C6h1206+602 >6h20+6CO2+36Atp
Cellular respiration
Which organisms did you test?
- What did you measure in order to determine the rate of respiration in these organisms? How were the rates calculated?
Germinating peas and super worms
Know how to interpret an absorbance spectrum. Which colors of light are the best /worst for photosynthesis and why?
Plants absorbs light in the red ad blue regions during photosynthesis
These colors correspond to the wavelengths that chlorophyll pigments can absorb mot efficiently.
Green light is only reflected and gives the plants its color. The plant will bot get light to grow ad carry out photosynthesis
What is the overall equation for photosynthesis? How do photosynthesis and cellular respiration relate to each other?
6CO2+6H2O+LIGHT ENEGRY> C6H12O2+6O2
Photosynthesis provides the oxygen and glucose needed for cellular respiration while cellular respiration produces the carbon dioxide and water needed for photosynthesis.
How does agarose gel electrophoresis work?
It is used to separate DNA fragments bases on their size and charge.
The gel acts as a sieve allowing smaller molecules to pass through more easily than larger ones.
What is a restriction enzyme? How do RFLPs and restriction enzymes relate to each other? Finish answering
It is a endonucleases which catalyze the cleavage of phospdiester bonds within both strands of DNA.
They relate to each other because
What is RFLP? Why is it used?
Restriction FRAGMENT LENGTH POLYMORPHISMS
It is sued to manifest the unique molecular genetic profile or fingerprint of an individuals dna.
