Test 2, 15. analysis of cells, molecules, and systems

Question 1

What is a cell culture?

Answer 1

1. removing cell form organism and promoting their growth in a favorable artificial envrionment

Question 2

What is a primary cell culture?

Answer 2

1. derived from animal directly
2. survive for finite time period
3. mechanical tissue disruption and isolation of tissue and cells to obtain sample

Question 3

What is a continuous cell line?

Answer 3

1. line of primary cells that have become immortal bc of transformation
2. tumor derived, or from viral transformation (epstein barr)

Question 4

Primary cell cultures are finite, and can only divide a number of times. This is senescence or________?

Answer 4

1. the loss of the ability to proliferate, based on genetic

Question 5

What are characteristics of cell line cultures>

Answer 5

1. more differentiated phenotype
2. homogenous population
3. infinite in vitro life span
4. immortalized from spontaneous genetic mutation

Question 6

What are some examples of cell line cultures?

Answer 6

1. SH-SY5Y

2. parkinson’s model of disease

Question 7

What morphological categories are cell lines divided into?

Answer 7

1. fibroblastic
2. epithelial like cells
3. lymphoblast like cells

Question 8

What are fibroblastic cells?

Answer 8

1. mammalian cell line
2. multipolar/bipolar
3. elongated shape that grows onto substrate

Question 9

What are epithelial-like cells?

Answer 9

1. mammalian cell line
2. polygonal shape
3. attach to substrate in discrete patches

Question 10

What are lymphoblast like cells?

Answer 10

1. mammalian cell line
2. spherical shape
3. grow in suspension, not surface attachment

Question 11

What are the advantages of cell cultures?

Answer 11

1. observe cell behavior without animal variations
2. maintain cells for generation= reproducibility
3. controlled growth environment
4. exposure to specific isolated reagents

Question 12

What are the disadvantages to cell culture growth?

Answer 12

1. require standardized technique
2. time consuming
3. limited material
4. loss of original cellular mechanisms

Question 13

Where can cell cultures be applied to life and sciences?

Answer 13

1. cell/gene function research
2. biological product formation
3. testing
4. regenerative medicine

Question 14

Why is protein purification important?

Answer 14

1. study structure and function of specific protein

Question 15

What is recombinant DNA technology used for?

Answer 15

overexpress a specific protein making it easier to purify

Question 16

What are the steps of sub-cellular fractionation?

Answer 16

1. mechanically blend tissue, to form homogenous mix
2. centrifuge
3. cell lysis
4. ultracentrifugation

Question 17

What techniques can be used to lyse cells?

Answer 17

1. osmotic shock
2. ultrasonic vibration
3. mechanical blending

Question 18

What is ultracentrifugation used for?

Answer 18

1. separate organelles

Question 19

What are lipid rafts?

Answer 19

1. PM domains high in cholesterol, sphingolipids, gangliosides
2. detergent insoluble
3. center for signal transduction
4. abnormal protein processing in neurodegenerative disorders

Question 20

What are the different matrices used for column chromatography?

Answer 20

1. ion-exchange
2. gel-filtration
3. affinity

Question 21

What is ion-exchange chromatography?

Answer 21

separation of compounds based on their attraction to the column based on ion charge.

Question 22

What is gel-filtration chromatography?

Answer 22

separation of compounds based on size. Column has certain pore size, that compounds can get trapped in and retained. Larger particles drain faster, smaller drain more slowly

Question 23

What is affinity chromatography?

Answer 23

similar to ion-exchange except charge is not used. Use the affinity of a substrate to an exnzyme. Stronger affinity equals longer retention time

Question 24

How are genetically engineered proteins formed?

Answer 24

1. use of recombinant DNA technology

2. use tags to purify the protein and recognize specific epitopes

Question 25

What is SDS used to do, when analyzing proteins in an SDS-PAGE experiment?

Answer 25

1. unfolds the proteins 2. attaches uniform negative charge 3. contains beta-mercaptoethanol to reduce disulfide bonds

Question 26

What feature of proteins does SDS-PAGE use to evaluate different proteins in a solution?

Answer 26

1. the size of the denatured protein

Question 27

What feature is critical to the analysis of proteins when using a 2-dimensional gel?

Answer 27

1. isoelectric points and the pH of the solution

Question 28

If the pH is higher than the isoelectric point of a compound, which direction will it move?

Answer 28

the protein will migrate towards the + pole

Question 29

If the pH is lower than the isoelectric point, which direction will the compound migrate?

Answer 29

the protein will migrate towards the - pole

Question 30

If the pH is equal to the isoelectric point, which direction will the compound move?

Answer 30

it will not move.

Question 31

What is Sirt3 and what happens if it malfunctions?

Answer 31

1. mitochondrial protein deacetylase | 2. malfunciton= increased acetylated Sirt3 substrates

Question 32

What is Western Blotting?

Answer 32

1. type of protein analysis that uses Ab-Ag interaction in order to determine a protein.

Question 33

What is indirect immuno-cytochemistry?

Answer 33

1. type of western blotting that utilizes secondary Ab that are marked and will bind with a primary Ab-Ag interaction. 2. very sensitive method

Question 34

How is ELISA useful in protein analysis?

Answer 34

1. provides protein detection and quantification

Question 35

How is ELISA used to determine presence of Ag?

Answer 35

1. Ab-enzyme complex binds with the Ag, forming a color and absorbance level is then analyzed with machine

Question 36

When would Indirect ELISA be used?

Answer 36

1. used to detect the presence of an Ab 2. used for HIV infection 3. use enzyme linked Ab react with Ag

Question 37

What does an enzyme reaction suggest for an indirect ELISA test?

Answer 37

1. enzyme-linked antibodies were bound to human antibodies and implies the Pt has antibodies to viral antigen

Question 38

What is sandwich ELISA?

Answer 38

1. detects the antigen and the quantity of Ag. | 2. uses standard curve in order to determine the amount of Ag present