Brainscape's Knowledge GenomeTM


Top Flashcards (Certified)
All Flashcards

chem semester 2 > electrophoresis > Flashcards

electrophoresis Flashcards

1
Q

What are the two kinds of electrophoresis?

A
  1. Zone electrophoresis (majority)
  2. Moving Boundary Electrophoresis
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What is Zone electrophoresis?

A

Moving proteins through a solid medium/matrix.

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

What are the types of Zone electrophoresis?

A
  1. Paper electrophoresis (cellulose acetate)
  2. Gel electrophoresis (agarose or polyacrylamide gel)
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What is Moving Boundary Electrophoresis?

A

Moving proteins through an aqueous type of medium.

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

What are the types of Moving Boundary Electrophoresis?

A
  1. Capillary electrophoresis
  2. Immuno electrophoresis
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What are the components of electrophoresis?

A
  1. Driving force - power supply
  2. Support medium - agarose gel, etc.
  3. Buffer
  4. Sample
  5. Detection System
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What role does the buffer play in electrophoresis?

A

Serves as an important component; connects the sample and electrical force, dictates pH.

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

What happens to proteins at different pH levels?

A

pH > pI: Negative charge
pH < pI: Positive charge

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

What is the goal of electrophoresis?

A

To ensure all proteins have the same charge.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What is the charge state of most proteins?

A

Most are isoelectric: neutral and will not be moving.

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

Why do negatively charged proteins absorb less water?

A

Negatively charged proteins absorb less water and buffer than positively charged proteins.

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

Which direction do serum proteins migrate?

A

Serum proteins migrate toward the anode, starting at the cathode.

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

What factors affect the movement of proteins?

A
  1. Weight of protein
  2. Velocity (depends on charge and strength of electrical field)
  3. Medium used
  4. Temperature
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

What is the maximum temperature for maintaining proteins during electrophoresis?

A

Heat cannot be above 55 degrees; will denature proteins.

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

What is the importance of ionic strength in electrophoresis?

A

Ionic strength directs migration; too low enhances movement, too high makes it sluggish.

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

What is the normal ionic strength range for electrophoresis?

A

Normal strength = 0.01 - 0.1

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

What materials are used for electrophoresis?

A

Proteins easily migrate in cellulose acetate and/or agarose gel.

maintain even temp

do not absorb water or buffer from system

prevent endo electrical osmosis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

What is the advantage of polyacrylamide gel?

A

More separation of proteins; separates based on molecular weight.

called poly because of diff layers of agarose gel

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

What is a common specimen used in electrophoresis?

A

Serum, diluted in buffer.

can also use urine /CSF
-no dilution; concentrate then

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

What is the typical volume of specimen used in electrophoresis?

A

Use 2-5 mL of specimen.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

What should be done before running electrophoresis?

A

Plug in the power supply before putting proteins in (like warm up the oven.)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

What are common detection methods in electrophoresis?

A

Stains such as ponceau (cellulose acetate) and coomassie blue (any agarose gel).

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

What is bromocresol purple used for?

A

Used in place of coomassie blue at pH 5.2 (proteins positively charged).

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

What is bromocresol green/yellow dye used for?

A

Used for detection of albumin.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
what is endo electrical osmosis
backward migration of proteins from starting ??
26
proteins migrate better if they have
negative charge
27
order of bands from cathode to anode
gamma B1 and B2 A2 A1 Albumin
28
normal total serum protein value
6-8 gm/ dL
29
normal AG ratio
0.8-1.8
30
what is the highest peak in electrophoresis
albumin
31
what is 1st protein lost in kidney disfunction
albumin
32
2 primary roles of albumin
1. helps maintain osmotic pressure 2. transport protein
33
reasons for decreases in albumin
liver dysfunction- albumin made here kidney disfunction inflammatory/ infectious processes - making more APR
34
reason for increase in albumin
dehydration
35
reason for bisalbuminemia (teak peaks)
hereditary, greek culture, drugs non pathologic event
36
main protein seen in alpha 1 region
alpha 1 anti-trypsin
37
purpose of alpha 1 anti-trypsin
Protein that will help with lung function; inhibits some protease that can be seen in certain diseases can be APR
38
decreases in alpha 1 anti-trypsin are
asthma, etc problems with lungs
39
other protein in alpha 1 region
alpha 1 fetoprotein -low levels after birth?? -tumor marker
40
what is alpha 1 region home to
APR- increases during infections
41
alpha 2 region main protein
haptoglobin - first thing measured in hemolytic event
42
another protein found in alpha 2
ceruloplasmin- binds copper
43
increased levels of copper shown in
wilson's disease - absorption not regulated decreased ceruloplasm
44
will not get lost in kidney: too large to pass
alpha 2 macro globulin -protective to fetus with mother's immune response
45
beta region protein
transferrin - binds 2 atoms of iron
46
iron overload and muscle damage; hemochromatosis = decrease
transferrin (all iron bound)
47
inflammation and IDA = increase
transferrin
48
another protein in beta region
CRP- c reactive protein APR
49
CRP only increases in
acute inflammatory (now)
50
gamma globulins made in
gamm region with plasma cells
51
normal or decreases albumin -increase A1 and A2; APR
acute inflammatory response
52
- Normal or decreased albumin - Increase A1 or A2 region - Increase in gamma region - Sometimes beta
chronic inflammatory response
53
- Plateau in the gamma region - Over production of antibodies??; auto immune diseases - Not a cancerous event; it’s a normal event May be transient
polyclonal gammopathy
54
- M spike or church spike/ peak - Can be higher than albumin - Be toward the cathode end ; gamma - Cancerous event - Overproduce 1 type of immunoglobulin ○ Non functioning antibodies from oncogene ○ Ex. Multiple myeloma - See presence of Bence jones proteins: free light chains
monoclonal gammopathy
55
hypogammaglobulinemia
not making antibodies B cell defect give them IV gamma globulins
56
characteristic of alcohol cirrhosis of the liver
beta- gamma bridge regions blur together GGT and osmol gap increased
57
SPE
serum protein electrophoresis
58
SPE is based on 2 principles
1) electrophoresis 2) precipitation
59
steps serum protein electrophoresis
1) ID presence of monoclonal gammopathies or polyclonal gammopathies or normal look for increase IgG, IgA, IgM, kappa, and lambada
60
specimen used for SPE
serum and urine done with agarose gel
61
how to perform SPE?
specimen placed cathode 2-5 allow electrophoresis 15-30 min once migrate, add anti-serum to see if precipitation band; incubate detection: add stain to read
62
polyclonal apperance on test
smudging
63
monoclonal appearance on test
distinct line
64
what do you look for in urine electrophoresis?
bence jones proteins -over production of free light chains
65
in liver disease graph
everything decreases besides gamma because not produced there
66
kidney disease graph
alpha 2 increases; macroglobulin -can't pass kidney
67
inflammatory graph
APR increases albumin normal or slightly decreased
68
spinal fluid normal protein
15-45
69
measure of nutrition in CSF electrophoresis
prealbumin
70
how many bands in normal CSF
7 -beta regions more distinct
71
hemorrhaging in CSF graph
increase in B2 because increase in fibrinogen
72
HGB alkaline used as a
screen
73
if something abnormal in alkaline use
acid

chem semester 2 (9 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2025 Bold Learning Solutions. Terms and Conditions