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CC LEC > Electrophoresis > Flashcards

Electrophoresis Flashcards

1
Q

→ A molecule that contains both acidic & basic groups (amino acid = carboxyl & amine groups)

A

AMPHOLYTE

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2
Q

→ Rate of migration of a charged solute in an electric field

A

ELECTROPHORETIC MOBILITY

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3
Q

→ Preferential movement of water in 1 direction through electrophoresis medium due to selective binding of 1 type of charge on the surface of the medium

A

ENDOSMOSIS

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4
Q

a record that consists of the separated components of a mixture produced by electrophoresis in a supporting medium

A

ELECTROPHORETOGRAM

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5
Q

: type of electrophoresis limited to migration of small ions

A

IONTOPHORESIS

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6
Q

: type of electrophoresis usually used for migration of macromolecules

A

ZONE ELECTROPHORESIS

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7
Q

The movement of electrically charged compounds in a medium resulting to their separations based on their electrical charges when an electric current is applied

A

Electrophoresis

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8
Q

Macromolecules found in:

A

-proteins in serum
-urine
-cerebrospinal fluid (CSF)
-erythrocytes and tissue and,
-other biologic body fluids

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9
Q

ELECTRODES:

A

a. Cathode —- negatively charged

b. Anode —- positively charged

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10
Q

Factors Affecting Rate of Migration

A

Net electric charge of the molecule
Size and shape of the molecule
Strength of electrical field
Temperature of operation
Properties of Supporting Medium

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11
Q

Components of Electrophoresis

A
  1. Driving force (electrical power)
  2. Support Medium
  3. Buffer
  4. Sample
  5. Detecting System
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12
Q

In electrophoresis, heat is produced when_____ flows through a medium that has_____, resulting in an increase in thermal agitation of the dissolved solute (ions) and leading to a decrease in resistance and an increase in current.

A

Driving force (Electrical Power)

current; resistance

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13
Q

Supporting Medium

A

Cellulose acetate
Agarose gel
Polyacrylamide gel

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14
Q

Cellulose is acetylated to form cellulose acetate by treating it with ______

a dry, brittle film composed of about___ air space

homogenous medium with uniform pore size and does not absorb the protein

A

Cellulose acetate

acetic anhydride; 80%

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15
Q

A highly purified uncharged polysaccharide derived from agar requires small amounts of sample (approximately 2 mL) it does not bind protein and, therefore, migration is not affected

A

Agarose gel

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16
Q

referred to as ____

layers of gel with different pore sizes are used

separates serum proteins into___ or more fractions rather than the usual five fractions separated by cellulose acetate or agarose

A

Polyacrylamide gel; PAGE

20

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17
Q

-separates proteins on the basis of surface charge and molecular size, as does polyacrylamide gel.

-not widely used because of technical difficulty in preparing the gel

A

Starch

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18
Q

o Polysaccharide extracted from sea weed.
o Gel casted horizontally

A

Agarose

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19
Q

o Non-toxic.
o Separate large molecules

A

Agarose

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20
Q

o Commonly used for DNA separations.
o Staining can be done before or pouring the gel.

A

Agarose

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21
Q

o Cross-linked polymer of acrylamide.
o Gel casted vertically.

A

Polyacrylamide Gel

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22
Q

o Potent neuro-toxic.
o Separate small molecules.

A

Polyacrylamide Gel

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23
Q

o Used for DNA or protein separations.
o Staining can be done after pouring the gel.

A

Polyacrylamide Gel

24
Q

is a molecule whose net charge can be either positive or negative

A

AMPHOLYTE

25
Q

If buffer is more acidic than the pI -becomes_____ charged and migrates toward the_____

A

positively ; cathode

26
Q

If buffer is more basic than the pI -becomes____ charged and migrates toward the____

A

negatively

anode

27
Q

5 Buffers

A

Barbitone buffer - (around 8.0 pH)
Phosphate buffer -( around 7.0 pH)
Tris - borate - EDTA buffer (TBE)-(pH around 8.0)
Tris - acetate - EDTA buffer (TAE)- (pH around 8.0)
Tris - glycine buffer - (pH more than 8.0)

28
Q

• serum protein separation,
• poor resolution, weak buffer.

A

Barbitone buffer - (around 8.0 pH)

29
Q

• Enzyme separation,
• low buffering capacity.- high conductivity

A

Phosphate buffer-( around 7.0 pH)

30
Q

• Nucleic acid Separation
• Good resolution , high buffering capacity , low conductivity.

A

Tris - borate - EDTA buffer (TBE) -(pH around 8.0)

31
Q

• Nucleic acid separation
• high resolution, high buffering capacity low conductivity.

A

Tris - acetate - EDTA buffer (TAE)- (pH around 8.0)

32
Q

• Protein separation
• high buffering capacity , low conductivity

A

Tris - glycine buffer - (pH more than 8.0)-

33
Q

Stains

PROTEINS

A

• Amido Black
• Coomassie Brilliant blue
• Bromphenol Blue

34
Q

Stains

DNA

A

• Ethidium Bromide
• Sybr Green or Sybr Gold

35
Q

Stains

LIPOPROTEINS

A

• Sudan Black

36
Q

HEMOGLOBINS

A

• Ponceau Red
• Amido Black
•Coomassie Briliant blue

37
Q

It measures the absorbance of stain- concentration of the dye and protein fraction.

A

Densitometry

38
Q

Densitometry

BASIC COMPONENT:

A

1.light source
2.monochromator
3.movable carriage
4.photodetector
5.read-out device

39
Q

•α1-fetoprotein
•α-antitrypsin
•HDL

A

α1-Globulins

40
Q

•haptoglobin
•ceruloplasmin
•α2-macroglobulin

A

α2-Globulins

41
Q

•transferrin
•C-reactive protein

A

β-Globulins

42
Q

•immunoglobulins

A

γ-Globulins

43
Q

based on refractivity (the ability of the substance to bend light)

A

Refractometry

44
Q

when the light passes from one medium into another, the path of the light beam changes direction at the boundary surface if its speed in the second medium is different from that in the first

A

Refractometry

45
Q

Parts of the Refractometer

A
  • Polarizing Filter
  • Glass Hemicylincer
  • Viewing Lens
  • Cover
  • Light Enters in Back
  • Metal Case
46
Q

homogenous medium with uniform pore size and does not absorb the protein

A

Cellulose acetate

47
Q

Agarose gel

A highly purified uncharged_____ derived from agar

A

polysaccharide

48
Q

Agarose gel

requires small amounts of sample (approximately____)

A

2 mL

49
Q

Supporting medium that has uniform pore size

A

Cellulose acetate

50
Q

Supporting medium that has different pore size

A

Polyacrylamide gel

51
Q

How many fractions are separated by cellulose or agarose?

A

5

52
Q

It scans and quantitate electrophoretic pattern.

A

Densitometry

53
Q

-the ratio of the two speed of light

A

Refractive Index

54
Q

Black wire

A

Negative

55
Q

Red wire

A

Negative

56
Q

Contains the buffet/ electrolyte solution

A

Subcell

57
Q

Where the sample went the sample is placed (usually near cathode)

A

Sample well

CC LEC (39 decks)

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