Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

AMM > IF, IHC and ICC > Flashcards

IF, IHC and ICC Flashcards

1
Q

What does IF stand for?

A

Immunofluorescence

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What does IHC stand for?

A

Immunohistochemistry

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What does ICC stand for?

A

Immunocytochemistry

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Where is the variable region of an antibody? Top or bottom?

A

Top

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What does the antibody bind to?

A

Epitope on the antigen

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

How many types of mammalian immunoglobulin chains are there?

A

5
IgA, IgD, IgE, IgG and IgM

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What are the three sections an antibody can be divided into?

A

Two Fab regions
One Fc region

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Which region of the antibody is the variable domain in?

A

Fab regions

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What binds to the Fc fragment?

A
  • Fc receptors on lymphocytes
  • Secondary antibodies
  • Dye labels and enzymes
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What enzyme do we use if we want to produce a fragment both Fab regions and no Fc?

A

Pepsin

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What enzyme do we use if we want to produce all three parts?

A

Papain

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Which antibody is widely produced, found in mucosal areas and prevents colonisation of pathogens?

A

IgA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Which antibody is found with IgM on the surface of B cells? And is involved in B cell development?

A

IgD

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Which antibody is important in allergies and binds to allergens causing histamine release?

A

IgE

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Which is the major antibody and produces the most antibody-based immunity against invading pathogens?

A

IgG

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Which is the first response antibody, has high avidity (strength) and eliminates pathogens in early B cell immunity? (before sufficient IgG?)

A

IgM

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

Which antibody type is a mixture of many types of antibodies?

A

Polyclonal antibody

18
Q

Why are multiple antibodies produced in response to an antigen?

A

Because antigens have multiple epitopes.

19
Q

From where in the animal’s body are antibodies collected?

A

Antiserum (whole serum following exposure to antigen).

20
Q

Why are lab animals injected twice with the same antigen?

A

2nd infection activates memory cells to make IgG antibodies.

21
Q

What is affinity maturation?

A

Affinity maturation is the process by which B cells undergo selection to produce antibodies with increased binding affinity for a specific antigen.

Leads to a more effective immune response upon re-exposure.

22
Q

What is an adjuvant?

A

Adjuvant is a substance that is added to a vaccine to enhance the body’s immune response to the vaccine’s antigen.

Stimulates greater antibody production

23
Q

Monoclonal antibody production:
What is collected from the lab animal after infecting it? And where in the body it collected from?

A

B cells
Collected from spleen of lab animal

24
Q

What are collected B cells fused with in monoclonal antibody production?

A

B cells fused with immortal cancerous B cells (myeloma cells).

25
Q

Why are B cells fused with immortal cancerous B cells?

A

To yield hybridoma cells that can proliferate and produce antibodies continuously.

26
Q

What are hybridoma cells screened for?

A

Desired monoclonal antibody (recognising single epitope).

27
Q

How do we develop a secondary antibody?

A

Immunise a second animal with IgG from mouse.

Animal produces IgG in response to constant region of mouse IgG.

28
Q

What is immunofluorescence?

A

Visualises presence and location of specific proteins/ antigens in cells/tissues using antibodies labeled with fluorescent dyes

29
Q

Can Immunofluorescence also be used to detect protein modifications?

A

Yes

30
Q

What are the two types of Immunofluorescence (IF)?

A
  1. Direct
  2. Indirect
31
Q

What is the purpose of the fixation step in Immunofluorescence (IF)?

A

To preserve original cell architecture and ensuring antigens are immobilised.

32
Q

What is the purpose of organic solvents such as methanol and acetone in the fixation step of IF?

A

Organic solvents remove lipids from the cell membrane, facilitating antibody binding.
Acetone is more harsh.

33
Q

What types of aldehyde are used in aldehyde based fixing?

What is the purpose of aldehyde-based fixing?

A

4% formaldehyde or 10% formalin

To cross-link proteins.

34
Q

Why is permeabilisation step in IF needed?

A

Allows antibodies to access intracellular antigens within cells.

Creates openings in membrane.

35
Q

If we used organic solvent in fixation step, do we need to permeate?

A

No, as cell membrane holes are already punched.

36
Q

If aldehyde fixative used, do we permeate?
If so, what do we use to do so?

A

Yes.
Non-ionic detergent such as Triton X-100 or saponin.

37
Q

How do we reduce non-specific antibody binding during IF?

A

Blocking

38
Q

What are two commonly used blocking reagents in IF?

A

1% milk powder in PBS
Serum from the 2nd animal

Both bind unoccupied sites

39
Q

What is the function of the microtone during IF and IHC?

A

To create uniform sections of tissue samples of a specified thickness.

40
Q

How does ImmunoHistoChemistry (IHC) differ from Immunofluorescence (IF)?

A

secondary antibody is enzyme-linked in IHC

41
Q

How does ImmunoCytoChemsitry (ICC) differ from ImmunoHistoChemsitry (IHC)?

A

ICC involves staining cells (with secondary antibodies), not tissue.

AMM (18 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions