labs Flashcards
what kind of electrophoresis are we using?
polyacrylamide gel electrophoresis (PAGE)
what is PAGE used for
separating small molecules such as proteins
how many common amino acids
20
primary protein structure
sequence of amino acids in the chain
secondary protein structures
H bonding between components of backbone (helices, sheets)
where are disulfide bonds and H-bonding between side chains found
tertiary and quaternary structures
tertiary structure
interaction of hydrophilic and hydrophobic side chains with aqueous environment
ex. of tertiary. proteins
structural and globular proteins
quaternary structure
interaction of polypeptide chains with others
ex. of quaternary proteins
- myosin
- hemoglobin
what was PAGE analysis first carried out for
showing sickle cell anemia caused by change to single amino acid of hemoglobin
what detergent are we using
sodium dodecyl sulfate (SDS)
is SDS anionic or cationic
anionic
function of SDS and heat
- denatures the protein tertiary and quaternary structures so the proteins become more linear
- SDS gives protein an overall negative charge with strength that is relative to length of chain
where do the proteins flow from
negative cathode to positive anode
do larger or smaller proteins move more slowly
larger
how are proteins measured
Daltons (mass of hydrogen atom - 1.66 x 10^-24 g)
- use kilodaltons
average weight of amino acid
110 daltons
what is the tank buffer called
1x TGS
components of tank buffer
- tris
- glycine
- SDS
tris
modulates pH and Cl-
glycine
provides important ion source
sample buffer name
Laemmli
Laemmli sample buffer components
- tris
- SDS
- bromophenol blue
- beta-mercaptoethanol
- glycerol
bromophenol blue
relative location of our sample
beta-mercaptoethanol
reduces disulfide bonds
glycerol
increases density of sample so it falls into well
boiling breaks all but what?
disulfide
fixative ratio
50:10:40 ethanol:acetic acid: water
purpose of fixative
- removes SDS for better staining
- fixes proteins in place in gel
approx weight of protein
aa x 110 daltons
why 15 times
dislodges the membrane just enough to get the proteins out
contractile elements of muscle cells
myofibrils that are bundled into muscle fibers
each myofibril consists of contractile units called __________
sarcomeres
when does the sarcomere shorten
when myosin hydrolyzes ATP
biggest additional protein used in muscle contraction
titin
titin function
centers myosin in sarcomere
tropomyosin function
strengthens actin filaments
myosin heavy chain
slides actin filaments
myosin light chain
slides actin filaments
what website did we use
fishbase.net
what does an evolutionary tree/cladogram show
evolutionary lineages of different species over relative time
the Gnathostoma fishes are divided into which classes?
- Chondrichthyes
- Osteichthyes
2 subclasses of Osteichthyes
- Sarcopterygians
- actinopterygians
Sarcopterygians
living fossil, coelacanth, tetrapods
actinopterygian
modern ray-finned fish
