BIO 169 Lab
Enzyme Linked Immunosorbant Assay
What is ELISA used for?
used as an initial test for HIV detection. Also, influenza virus, bacterium that causes Lyme Disease, smallpox virus, SARS coronavirus, West Nile and others.
What are the two types of ELISA
Direct and Indirect
Indirect ELISA tests for?
patients blood for presence or absence of antibodies against a particular pathogen
What does the presence of antibodies indicate?
the individual has been infected and their body has produced an immune response.
Direct ELISA tests for?
the presence of antigens in the patients blood
What is ELISA based on?
the principle that antibodies produced in response to pathogens attach to their antigen targets specifically to form antigen-antibody complexes
Steps of ELISA
1. antigen proteins purified from the infectious agent have been added to and been bound to the bottom of the wells
2. Blood serum from each patient is added to the treated wells. If the serum contains antibodies against the bound antigen the antibodies will attach, forming tight complexes
3. Wells are washed between each step to remove any unbound proteins (visualization is the next step).
4. A second antibody is added that recognizes antibodies produced by humans
5. Chromogen substrate added to well. If present, the enzyme will facilitate a chemical reaction that changes the color of the chromogen
6. Color change indicates patient has been infected.
The catabolic process that breaks down large food molecules to monomers?
Break down of substances into simplier forms
chemical building blocks
breakdown of any food molecule by enzymes
Hydro - water
lysis - broken
Common Carbohydrates in our diet are
glucose, fructose, galactose, sucrose, lactose, maltose, glycogen and starch
the intestines can only absorb what type of carbohydrate
What form are most carbohydrates in?
enzyme in sylvia that spilts starch into oligosaccharides?
What environment does amylase work best in?
slightly acidic to neutral
Stomach acid inactivates?
Pancreatic amylase released by the pancreas into the small intestines finishes?
What are the most important brush border enzymes?
dextrinase and glucoamylase
Lactose intolerant is
the inability to digest milk sugar
Monomers of protein are:
Are we actually cannibals?
Where does protein digestion begin?
Pepsinogen is activated to form what protein digesting enzyme in the stomach?
What environment does pepsin work best in?
Acidic or basic?
What five enzymes in the small intestines continues protein digestion?
trypsin, chymotrypsin, carboxypeptidase, aminopeptidase and dipeptidase.
What are the most abundant fats in the diet?
Primary site for fat digestion is
what fat digesting enzyme does the pancreas produce?
The detergent (soap) like material allows the fat to mix with the watery environment of the body?
An emulsion is?
an aqueous suspension of fatty droplets
Pancreatic lipases cleave off two fatty acid chains, yeilding free:
fatty acids and monoglycerides