quiz 6

Question 1

cytochemistry

Answer 1

staining individual cells

Question 2

histochemistry

Answer 2

staining thin sections of tissue

Question 3

a technique that takes advantage of the highly specific interaction of antibodies with their target antigens to locate cellular molecules.

Answer 3

Immunostaining

Question 4

Cells must therefore be preserved in some manner prior to
antibody treatment through process called

Answer 4

fixation

Question 5

why must cells be fixed before immunostaining?

Answer 5

A typical live cell taken through the processes necessary for immunostaining would undergo a significant amount of degradation by the time staining was completed

Question 6

________ permanently “freezes” cellular molecules in place by
using chemical reactions rather than low temperature

Answer 6

Fixation

Question 7

The tertiary structures of proteins are largely maintained by _______________

Answer 7

noncovalent interactions

Question 8

loss of 3D shape on protien

Answer 8

denaturation

Question 9

is a solution of the gas formaldehyde, which forms covalent chemical crosslinks between the different parts of protein molecules and prevents denaturation.

Answer 9

Formalin

Question 10

version of formalin called _________________ to fix Tetrahymena cells

Answer 10

paraformaldehyde

Question 11

___________are large protein molecules that cannot cross membranes, so cells are usually treated with a detergent or some other agent to create openings in the lipid bilayer that give antibodies access to internal antigens.

Answer 11

Antibodies

Question 12

permeabilization

Answer 12

removes more cellular membrane lipids to allow large molecules like antibodies to get inside the cell.

Question 13

Cells can be permeabilized using a mild detergent, such as Triton X-100 or NP-40.
T/F

Answer 13

T

Question 14

___ “soaks up” non-specific protein-binding sites, ensuring that the only
way the antibodies can bind is via interaction with their specific targets

Answer 14

BSA

Question 15

Why are antibodies produced?

Answer 15

Antibodies are produced in response to the presence of an agent that the immune system recognizes as being not part of the body.

Question 16

What is antibody structure

Answer 16

Antibodies are composed of four polypeptide chains (two light chains and two heavy chains) arranged like a capital letter “Y

Question 17

the portion of the molecule the antibody binds to is called an _____

Answer 17

epitope

Question 18

the molecule containing the epitope is referred to as an ______

Answer 18

antigen

Question 19

The specificity is so great that _________ can be generated to
distinguish between two versions of the same protein, one of which has a particular functional group attached and another that does not.

Answer 19

antibodies

Question 20

high level of _______ makes antibodies
extremely useful for locating and identifying
proteins or other antigens, and is employed in
the diagnosis of disease agents, detection of
hormones, proteins, or drugs in the blood,
immunoblotting, immunohistochemistry, and
immunofluorescence.

Answer 20

specificity

Question 21

we will be using _________ immunofluorescence to detect a protein antigen in Tetrahymena

Answer 21

indirect

Question 22

chemically joining a fluorescent dye directly to an antibody that was raised against an antigen you’re interested in.

Answer 22

direct immunofluorescence

Question 23

In direct immunofluorescence, the antibody, with its fluorescent label, will bind to the target antigen in cells, causing it to _______ under a fluorescence microscope

Answer 23

“light up”

Question 24

that a different labeled antibody has to be prepared for every
antigen a laboratory is interested in studying. In addition, the sensitivity of the technique is relatively low since there is only one labeled antibody molecule bound to each epitope

Answer 24

problem with direct immunofluorescence

Question 25

the primary antibody that binds specifically to the target antigen is left unlabeled. It is then detected after it binds by using a fluorescently labeled secondary antibody.

Answer 25

indirect immunofluorescence

Question 26

multiple labeled secondary antibodies can bind to a single primary antibody molecule, the signal strength is “amplified,” resulting in greater _______

Answer 26

sensitivity

Question 27

In ___________ antibodies (Ab) are used to label specific structures
of a cell or tissue with a visible tag.

Answer 27

immunostaining

Question 28

2 antibodies used in this type.
1. Primary antibody detects an antigen (Ag).
2. Secondary antibody carries the tag and binds to the primary antibody

Answer 28

Indirect immunofluorescence

Question 29

In this experiment, indirect antibody-labeling system will be
used to detect the protein __________ in cilia

Answer 29

a- tubulin

Question 30

__________a molecule (often a protein) that can be
recognized by an antibody

Answer 30

antigen

Question 31

• New antibodies can be developed by ________ a
  host animal with an antigen you are interested in.
  – The host animal mounts an immune response to the
  foreign protein.
  – Desired antibody is purified from the animal’s serum.

Answer 31

injecting

Question 32

In antibody nomenclature, what is the order

Answer 32

1. Host = the animal species that made the antibody
2. Antigen = what the antibody binds to
3. Tag = the fluorescent label (or enzyme) that produces signal

Question 33

in Goat-anti-mouse IgG-FITC, what is the host

Answer 33

goat

Question 34

in Goat-anti-mouse IgG-FITC what is the antigen

Answer 34

mouse IgG

Question 35

in Goat-anti-mouse IgG-FITC, what is the tag

Answer 35

FITC

Question 36

crosslinks proteins together to lock them in place

Answer 36

Fixation –

Question 37

– saturates non-specific binding sites with a generic protein

Answer 37

blocking

Question 38

Incubation with primary antibody – binds antigen

Answer 38

T

Question 39

what does Incubation with secondary antibody do

Answer 39

binds primary antibody