Techniques
Lab
Gel electrophoresIS:SEPARATE
·
charge
cathode
o
by the
and /or size
LARGE less movement
*
I
=
-
cannot
move as
well
-
*
small:BOTTOM
TOP
anoder
①
Large:
Samples in lanes
move toward Danode
~
Native-PAGE
for
·
separate
by
-
~
-
-
WITHOUT
separate
by mass
·
-
size
(keep structure)
SDS-PAGE for Proteins
denatures
-
Proteins
Reducing
breaks
breaking disulfide
(only noncovalent)
SDS-PAGE for Proteins
desulfide
COMPLETELY
bridges
denature
"SNOW DROP"
Western Blot PROTEINS
·
=
1. SDS
Page
conducted
2. Introduce radiolabeled antibodies
3. proteins bound to itshow up
Southern/Northern BLOT=DNA, RNA
·
FOR DNA
~
FOR RNA
deave/denature
s
~get electrophoresis
of single strands
polyacrylamide
*
agarose
*
get electrophoresis
single
of
strands
for <500 base pairs
for +500 base pairs
·
Chromotography:
separate the
components of
moble=nonpolar
*
muxture
stationary polar
*
Polar Slower)
-
=
-Liquid stationary
-
-
Gas-gas
en
Inert
silica, nonpolar mobile
mobile,
is
a
coating
is station.
(N2, heum)
-
Polar molecules elute SLOWER
longer retention times)
Size exclusion
-
-
larger
elute FASTER
smaller
L
-
SLOWER
proteins separated based
-
affinity for their ligand
HIGHER
affinity SLOWER
on
=
-
·TLC
solution is mobile, the
beads are station.
get
"caught"and elute
>Affinity
RF
-
will bind to the beads
Silica
destance
gel stationary
=
*
solvent line
and the
nonpolar mobile solvent
I nonpolar EasterI
more
Distillation:separate two
·
or more
molecules from solution
->
->
C
pts. 2 So apart
simple-boiling
fractional-boiling pts.
22 So apart
Durin
g
Glycogenesis
.
GLUT 2
Fed State
"GIP"
↑
~
Glucokinage
OR
Hexokinage
~
the
P
Mutage
·
UTP
iodgsine)
+
.
↓
ATP
*
6 Phosphorylate
1
Phosphogluco
·
on
the liver
10
&Gloose
receptors
UDP-gloose
auto
glucose
G
11
Keep
adch.
ingree
glycosylation
I
I
·
=
oD
*
driven
by
release of
dephos. from UTP
c.-
·
make
:
a
Branching
GLYOOGEN
1,6-glycosides Enzyme
SYNTHASE
GLYCOGEN
GLYCOGENIN
⑧o
Balboaahe
-
During
fasting
State
-
-
hypoglycera
Low blood
.
&
glucose
(glucagon, norepi/epi)
take 3
Slice
-,
glucose,
-
Enzyme
Diffuse out
cell
of the
little
1,0
1 GLUCOSE
cleave- 1,4
glycosidec
with phos.
very
glucose
for the
blood
·
R
make
Fore
Sfr
Debranching
*
make
i
-
1, 4 and transfer
to other, break
FORM
Glycogenolysis
*
S
&A
molecules
Phospho
gloco
mutage
↑ rose
ER
No
Glucose E Phosphatase
-
0
x
&
=
v
-100
sEs
g