D103 Review study Guide Exam
(Actual 2025/2026) Questions with
Complete Solutions | Latest Update |
Graded A+
Principles of immunofluorescence staining
we want to identify 2 dif proteins, how do we do it?
*Make sure that ........ (2 essential parts to note)✔✔First we must generate primary antibodies
that recognize/bind our target protein from 2 DIFFERENT species animals.
then, use secondary antibody (tagged with color) from a 3rd animal that recognizes/binds the
primary but is a dif color than the other secondary antibody
*primary antibodies must be from different species
*secondary antibodies must be a dif. color than each other, must be dif animal than primary
animal, but can be from same animal as other secondary
so basically, at least 3 dif animals required, refer to slide for better understanding
rabbit ;;;;;;;;;;;;;;;;;;;;;;;;;;;;mouse
goat ;;;;;;;;;;;;;;;;;;;;;;;;;;;;;;; goat
how and what is Tagging used for?✔✔through protein fusion of a fluorescent protein, we can
"tag" a protein of interest/target, making it fluorescent, so that we can see/monitor behavior
GFP✔✔green fluorescent protein, used for tagging, when shine blue light on it, reflects &
fluoresces green light back
Many types of tags and colors, name 2 examples please✔✔GFP: green fluorescent protein
RFP: red fluorescent protein
BFP: blue fluorescent protein, etc
,Immunofluorescense staining can be compared to a ___________ while Fluorescent tagging can
be compared to a ____________.✔✔-snapshot
-movie
Keep in mind that...✔✔a movie can be made of different snapshots so tagging can also be used
for immunofluorescent staining
another tagging technique used to observe protein dynamics is FRAP. what's that?✔✔-
Fluorescent Recovery After Photobleaching
-where they take a protein expressing GFP, and shoot a laser at a specific part, to determine how
fast the GFP signal/protein heals at that part, which shows how dynamic the protein is!
(similar to the scratch 'n' heal method)
Understand FRAP chart✔✔photobleaching disrupts GFP protein signal, as time passes measures
how fast the protein signal can be recovered to tell whether a protein is highly or low dynamic
fast recovery= high dynamics
slow recovery=low dynamics
btw dynamic= unstable (i think)
*the lower the bar the lower dynamic,
*the vertical line down represents 0 dynamics bc thats the photobleaching happening
Western blot assay✔✔technique that shows protein molecular weight
shows when the size changes either increase or decrease as compared to each row, must analyze
to find answer
refer to i-clicker question and practice
Cell sizes:
bacteria , yeast , oocyte , epithelial
largest? smallest? in between?✔✔largest: Oocyte
, smallest: bacteria
in between: yeast, epithelial,
major difference between prokaryotic and eukaryotic cells?✔✔eukaryotic contain organelles,
prokaryotic no organelles
to visualize/see small things we use a ____________. Some microscopes have limitations due to
_____________ and source ___________ capability.
If we want to see details about membrane, nuclear DNA, or an organelle, which type of
microscope do we use?
When we want to see immunofluorescence/tagging or regular cell shape, which type of
microscope do we use?✔✔-microscope
-light source , wavelength
-electron microscope (rly tiny- small wavelength)
-regular light microscope (regular- higher wavelength)
*wavelength is key
lipid dynamics: what factors affect membrane fluidity?✔✔1) length of fatty acid chain
-the shorter the FA chain, the less crowded and packed in it is, the more fluid the membrane
2) lvl of saturation of FA chain:
-more double bonds = less packed = more fluid
3) amount of cholesterol btwn lipids
-cholesterol acts as a rigid sticker, sticking FA's together instead of loose and personal space, so
the more cholesterol, the less fluid
Cholesterol✔✔-amphipathic membrane lipid
-can enhance hydrophobic region
-can increase membrane thickness which:
-"sticker" reduces membrane fluidity
Transmembrane Domain Graph
how to interpret?✔✔on table and graph:
(Actual 2025/2026) Questions with
Complete Solutions | Latest Update |
Graded A+
Principles of immunofluorescence staining
we want to identify 2 dif proteins, how do we do it?
*Make sure that ........ (2 essential parts to note)✔✔First we must generate primary antibodies
that recognize/bind our target protein from 2 DIFFERENT species animals.
then, use secondary antibody (tagged with color) from a 3rd animal that recognizes/binds the
primary but is a dif color than the other secondary antibody
*primary antibodies must be from different species
*secondary antibodies must be a dif. color than each other, must be dif animal than primary
animal, but can be from same animal as other secondary
so basically, at least 3 dif animals required, refer to slide for better understanding
rabbit ;;;;;;;;;;;;;;;;;;;;;;;;;;;;mouse
goat ;;;;;;;;;;;;;;;;;;;;;;;;;;;;;;; goat
how and what is Tagging used for?✔✔through protein fusion of a fluorescent protein, we can
"tag" a protein of interest/target, making it fluorescent, so that we can see/monitor behavior
GFP✔✔green fluorescent protein, used for tagging, when shine blue light on it, reflects &
fluoresces green light back
Many types of tags and colors, name 2 examples please✔✔GFP: green fluorescent protein
RFP: red fluorescent protein
BFP: blue fluorescent protein, etc
,Immunofluorescense staining can be compared to a ___________ while Fluorescent tagging can
be compared to a ____________.✔✔-snapshot
-movie
Keep in mind that...✔✔a movie can be made of different snapshots so tagging can also be used
for immunofluorescent staining
another tagging technique used to observe protein dynamics is FRAP. what's that?✔✔-
Fluorescent Recovery After Photobleaching
-where they take a protein expressing GFP, and shoot a laser at a specific part, to determine how
fast the GFP signal/protein heals at that part, which shows how dynamic the protein is!
(similar to the scratch 'n' heal method)
Understand FRAP chart✔✔photobleaching disrupts GFP protein signal, as time passes measures
how fast the protein signal can be recovered to tell whether a protein is highly or low dynamic
fast recovery= high dynamics
slow recovery=low dynamics
btw dynamic= unstable (i think)
*the lower the bar the lower dynamic,
*the vertical line down represents 0 dynamics bc thats the photobleaching happening
Western blot assay✔✔technique that shows protein molecular weight
shows when the size changes either increase or decrease as compared to each row, must analyze
to find answer
refer to i-clicker question and practice
Cell sizes:
bacteria , yeast , oocyte , epithelial
largest? smallest? in between?✔✔largest: Oocyte
, smallest: bacteria
in between: yeast, epithelial,
major difference between prokaryotic and eukaryotic cells?✔✔eukaryotic contain organelles,
prokaryotic no organelles
to visualize/see small things we use a ____________. Some microscopes have limitations due to
_____________ and source ___________ capability.
If we want to see details about membrane, nuclear DNA, or an organelle, which type of
microscope do we use?
When we want to see immunofluorescence/tagging or regular cell shape, which type of
microscope do we use?✔✔-microscope
-light source , wavelength
-electron microscope (rly tiny- small wavelength)
-regular light microscope (regular- higher wavelength)
*wavelength is key
lipid dynamics: what factors affect membrane fluidity?✔✔1) length of fatty acid chain
-the shorter the FA chain, the less crowded and packed in it is, the more fluid the membrane
2) lvl of saturation of FA chain:
-more double bonds = less packed = more fluid
3) amount of cholesterol btwn lipids
-cholesterol acts as a rigid sticker, sticking FA's together instead of loose and personal space, so
the more cholesterol, the less fluid
Cholesterol✔✔-amphipathic membrane lipid
-can enhance hydrophobic region
-can increase membrane thickness which:
-"sticker" reduces membrane fluidity
Transmembrane Domain Graph
how to interpret?✔✔on table and graph:
