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

Molecular BIO > Test 2, 15. analysis of cells, molecules, and systems > Flashcards

Flashcards in Test 2, 15. analysis of cells, molecules, and systems Deck (38):
1

What is a cell culture?

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

2

What is a primary cell culture?

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

3

What is a continuous cell line?

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

4

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

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

5

What are characteristics of cell line cultures>

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

6

What are some examples of cell line cultures?

1. SH-SY5Y
2. parkinson's model of disease

7

What morphological categories are cell lines divided into?

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

8

What are fibroblastic cells?

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

9

What are epithelial-like cells?

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

10

What are lymphoblast like cells?

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

11

What are the advantages of cell cultures?

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

12

What are the disadvantages to cell culture growth?

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

13

Where can cell cultures be applied to life and sciences?

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

14

Why is protein purification important?

1. study structure and function of specific protein

15

What is recombinant DNA technology used for?

overexpress a specific protein making it easier to purify

16

What are the steps of sub-cellular fractionation?

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

17

What techniques can be used to lyse cells?

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

18

What is ultracentrifugation used for?

1. separate organelles

19

What are lipid rafts?

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

20

What are the different matrices used for column chromatography?

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

21

What is ion-exchange chromatography?

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

22

What is gel-filtration chromatography?

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

23

What is affinity chromatography?

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

24

How are genetically engineered proteins formed?

1. use of recombinant DNA technology
2.use tags to purify the protein and recognize specific epitopes

25

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

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

26

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

1. the size of the denatured protein

27

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

1. isoelectric points and the pH of the solution

28

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

the protein will migrate towards the + pole

29

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

the protein will migrate towards the - pole

30

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

it will not move.

31

What is Sirt3 and what happens if it malfunctions?

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

32

What is Western Blotting?

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

33

What is indirect immuno-cytochemistry?

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

34

How is ELISA useful in protein analysis?

1. provides protein detection and quantification

35

How is ELISA used to determine presence of Ag?

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

36

When would Indirect ELISA be used?

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

37

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

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

38

What is sandwich ELISA?

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