Brainscape's Knowledge GenomeTM

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


See full index

Biochemistry semester 1 > Molecular separations > Flashcards

Molecular separations Flashcards

1
Q

reasons for biomolecule purification

A

study their properties
analyse distribution/ abundance
commercial/ medical use

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

synthetic biomolecules

A

incomplete reaction products resembling final product

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

2 main techniques for purification

A

electrophoresis
chromatography

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

relative centrifuge force

A

centrifugal force/ g force

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

microfuge

A

low speed

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

ultracentrifuge

A

high speed

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

isolation of protein sources in blood/ bacteria

A

centrifuge

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

isolation of protein sources in organs/ tissues

A

homogenisation

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

cell g force

A

3000g

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

organelle g force

A

20000g

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

protein g force

A

100000g

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

chromatography factors

A

size
charge
affinity

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

factors affecting affinity for stationary phase

A

charge interactions (ion exchange chromatography)
hydrophobicity (reverse phase/ hydrophobic interaction)
size (gel filtration/ size exclusion)
specific binding (affinity chromatography)

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

gel filtration

A

porous beads in which size affects distance travelled

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

ion exchange chromatography

A

elution via salt conc ^ competing for charge interactions
protein rich in charged residues

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

reverse phase chromatography

A

eluted by polarity reversal of solution via solvents

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

affinity chromatography

A

addition of free binding partner

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

immobilised metal affinity chromatography

A
  1. Histag addition
  2. elution
    *imidazole addition
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

histag

A

6-10 histidine residues at N/C terminus end
imidazole side chain specifically binding to metals

20
Q

addition of histag

A

addition of DNA coding for histag into natural DNA
bacteria produce protein

21
Q

histidine characteristic

A

heteroaromatic
- charged amine group

22
Q

histag elution

A

imidazole addition elutes
gradient/ step addition

23
Q

step elution

A

all bound molecules elute at once

24
Q

gradient elution

A

separates if different types bound

25
Q

gel electrophoresis gel function

A

convection current prevention so molecules move as a band, not dispersed

26
Q

gel pore size effect on gel electrophoresis

A

control macromolecule migration speed

27
Q

PAGE (polyacrylamide gel electrophoresis)

A

acrylamide polymers cross-linked w methylene bisacrylamide

28
Q

acrylamide polymer percentage effect

A

changes pore size

29
Q

pH of PAGE

A

> 8 therefore proteins are mostly negatively charged

30
Q

NATIVE-PAGE

A

Proteins separated by size, shape and charge

31
Q

colourless protein reveal in NATIVE-PAGE

A

blue-stain

32
Q

SDS

A

sodium dodecyl sulphate
*lauryl sulphate in shampoo

33
Q

SDS process

A

denatures proteins/ adds negative charge
shape/charge independent
heated w SDS/ mercapto-ethanol to disrupt disulfide bonds and denature

34
Q

SDS-PAGE

A

separation by protein size only
mercapto ethanol and heated to 95 degrees celcius denature
depends on pore size
marker proteins often used

35
Q

pore size proportion to molecular weight in SDS-PAGE

A

pore size proportional to log(MW)

36
Q

IEF

A

Iso-Electric Focussing

37
Q

IEF summary

A

Native-page carried out on pH gradient caused by ampholytes

38
Q

ampholytes

A

large polyions

39
Q

equilibrium technique IEF

A

proteins stop when they reach an isoelectric point

40
Q

what does IEF identify

A

proteins, their isomers and chemical modification

41
Q

2D Gel electrophoresis

A

IEF in tubular gels as first dimension, SDS page in second dimension

42
Q

isoelectric point/ pI

A

pH at which a particular molecule carries no net electrical charge

43
Q

what does 2D gel electrophoresis separate by?

A

size and pI

44
Q

DNA gel electrophoresis visualisation

A

UV light and fluorescent dyes

45
Q

DNA gel electrophoresis

A

small DNA fragments separated at high res via acrylamide gels in sequencing
larger fragements have larger pore gels
fixed negative charge density

Biochemistry semester 1 (7 decks)

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