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Module 04: Cellular Analysis Flashcards

(33 cards)

0
Q

What is done in mechanical lysing?

A

Rotating blades grind and disperse cells and tissues

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1
Q

___________ stop proteins and lipids from sticking together (solubilization).

A

Detergents

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2
Q

Once the cell is lysed and mixed into a homogenous solution, it is called:

A

Cell lysate (or cell homogenate)

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3
Q

centrifugation (n.)

A

the separation of particles depending on size or density

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4
Q

What is the unit of the sedimentation coefficient?

A

Svedberg units (S)

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5
Q

What does sedimentation coefficient measure?

A

The rate of particle sedimentation

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6
Q

Increasing sedimentation coefficient ________ the rate of sedimentation.

A

increases

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7
Q

What is relative centrifugal force (RCF) measured in?

A

xg, the relative multiple to the force of gravity

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8
Q

What is the equation for RCF?

A

RCF = 1.12 x R x (rpm/1000)²

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9
Q

What effect does doubling the radius have on RCF? (AEC)

A

Double the RCF

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10
Q

microsome (n.)

A

a vehicle derived from endomembrane system

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11
Q

polysome (n.)

A

a single mRNA with multiple ribosomes attaches, each synthesizing a polypeptide from the same mRNA

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12
Q

differential centrifugation (n.)

A

sequential centrifugation at higher so for longer times

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13
Q

Give a differential centrifugation sequence.

A

1000 xg for 10min; 20,000 xg from 20min; 80,000 xg for 1h; 200,000 xg for 2h

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14
Q

cellular homogenate (n.)

A

suspension of all material from broken cells

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15
Q

What do you get after the first round of DC?

A

Postnuclear supernant, and pellet containing nuclei and unbroken cells

16
Q

density gradient centrifugation (n.)

A

centrifuging using a concentration gradient (e.g., sucrose)

17
Q

Density of 2.3 M sucrose

18
Q

Density of lysosomes

19
Q

Density of mitochondria

20
Q

Density of peroxisomes

21
Q

What do you get after the second round of DC?

A

Postmitochondrial supernant, and pellet containing large particles (mitochondria, lysosomes, and peroxisomes)

22
Q

What do you get after the third round of DC?

A

Postmicrosomal supernant, and pellet containing fragments of cell membranes, smooth ER, and rough ER

23
Q

What do you get after the fourth round of DC?

A

Cytosol, and pellet containing viruses, free ribsosomes, and large macromolecules

24
What does coomassie stain?
Protein
25
What color does coomassie stain?
Brilliant blue
26
What does SDS do?
Denatures proteins
27
native state (n.)
folded and functional state of protein
28
What does DTT stand for?
Dithiothreitol
29
How many polypeptide chains comprise insulin?
Two polypeptide chains
30
What are the separate polypeptide chains insulin held together by?
Disulfide bonds
31
What kind of agent is DTT?
A reducing agent
32
What are the steps of a Western blot/immunoblot?
SDS polyacrylamide gel electrophoresis, protein blot on nitrocellulose, antibody labeling, antibody detection