FINAL review Flashcards
what is purpose of Lowry assay
determine protein level in solution
What color is generated in Lowry assay and what absorbance level is it measured
blue color; 750nm and 540 nm
the total protein concentration in Lowry assay is exhibited by what
color change of sample solution in proportion to protein concentration
how is absorbance measured
using spectrophotometer
Lowry reagent \/ Colin phenol is
a strong acid
in bradford assay, the absorbance is at
595 nm
what amino acids are required for Lowry assay
tyrosine and tryptophan
Lowry assay requires samples containing only
10-200 ug/ml of protein
Lowry assay generates ____ color; biuret assay results in _____ color
blue; purple
what two groups are found in an amino acid
amino group and carboxylic acid group
titration curve is a plot of what
pH of weak acid against degree of neutralization of acid by strong base
what does a zwitterion contain
a protonated amino group and deprotonated carboxyl group
what titrants are used in amino acid titration
1M KOH
how is unknown amino acid identified in amino acid titration
calculating its molecular mass
ninhydrin reaction is what type of reaction
redox reaction
what groups are involved in ninhydrin reaction
amino group of sample and ninhydrin reagent
what is end product formed similar to
di-dehydrin
di-ketohydrin has what color and what is it also known as
deep blue pigmentation; ruhemann’s purple
what is ninhydrin reagent used for
qualitative and quantitive colorimetric analysis for amino acids and sometimes for peptides and proteins
what molecules are used in part A of ninhydrin reaction
glucose, serine, and glycine
what technique is used and what is its purpose in ninhydrin reaction
thin layer chromatography; used to separate and identify amino acids
what result is a positive test in ninhydrin reaction
purple color
what is function of protease
enzyme that catalyzes proteolysis or breakdown of protein (break down peptide bonds)
how is protease activity measured
colormetric method
what are protease enzymes involved in
cell division, growth, apoptosis, and migration
TCA is used in protease activity to do what
stop the reaction by precipitating enzymes and unreacted substrates
ninhydrin reagents can be used in protease activity for what
quantification of TCA soluble peptides
what enzymes are used in protease activity
trypsin and substrate casein
enzyme kinetics is study of
rates of enzyme catalyzed chemical reactions
trypsin plays role of ___ and casein plays role of _____
enzyme; substrate
what molecule is used to stop enzyme catalyzed reaction
trichloroacetic acid
molecular weight of casein is
33,300 daltons
what is name of v vs [s] plot
Michaeles Menten plot
velocity is ____ variable and substrate conc. is ___ varible
dependent; independent
galactose is _____ because its aldehyde group is easily oxidized to carboxylic acid
reducing sugar
what result does sucrose give in resorcinol test
positive result
sucrose and trehalose are what type of sugars
non-reducing sugars
purpose of carbohydrate analysis experiment
test characteristics of sugars
Benedict’s reagents tests is for
reducing sugar
what is positive result in beneditcs reagent; Molisch test; resorcinol test; iodine-KI test
greenish red color (reducing sugar); red violet color (carbohydrate presence); red for ketone sugar and identifies between aldose and ketone sugar; blue color in presence of amylose/starch
what is purpose of DNA plasmids isolation and PCR
extract DNA plasmids from cell cultures and measure DNA concentration
what are steps for PCR
denaturation, annealing and extenstion
SDS has what purpose in DNA isolation lab
detergent that destabilizes cell wall and cause cell wall to burst and release concents
TE contains what
Tris-HCl which is a buffer to obtain correct pH
EDTA is what
chelates ions needed by DNAse, inactivate DNAses and keep DNA from degrading
another name for DNA fragment is
amplicon
DNA absorbs light at
260 nm and protein at 280nm
Agarose gel electrophoresis is good at separating fragments ranging from
100 bp to 25kb
purpose of DNA agarose gel lab
check PCR product and DNA size
what is DNA agarose gel made of
0.5 g agarose, TBE buffer, and ethidium bromide
what is purpose of using UV light in DNA agarose gel
visualize DNA
ethidium bromide bonded to DNA will glow what color at what wavelength
orange; 302
comparing mobility of DNA of known size to mobility of
your PCR fragment
purpose of gel filtration chromatography is
purification of biological macromolecules
gel filtration chromatography can separate protein of
different masses
how is molecular mass of unknown protein found
by comparing to elution volumes of molcular mass standards
how is hydrodynamic radius determined
by fluidity instruments …
AGE stands for
polyacrylamide gel electrophoresis
the unique migration rate of protein in SDS PAGE is found by
size of protein
SDS is considered what
an anionic deteregent
large negative charge of SDS can
impart once bounded to protein
what is SDS PAGE used for
separate molecules of DNA RNA or protein on basis of size or electric hcarge
