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

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

1
Q

What is a cell culture?

A
  1. removing cell form organism and promoting their growth in a favorable artificial envrionment
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2
Q

What is a primary cell culture?

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

What is a continuous cell line?

A
  1. line of primary cells that have become immortal bc of transformation
  2. tumor derived, or from viral transformation (epstein barr)
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4
Q

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

A
  1. the loss of the ability to proliferate, based on genetic
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5
Q

What are characteristics of cell line cultures>

A
  1. more differentiated phenotype
  2. homogenous population
  3. infinite in vitro life span
  4. immortalized from spontaneous genetic mutation
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6
Q

What are some examples of cell line cultures?

A
  1. SH-SY5Y

2. parkinson’s model of disease

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7
Q

What morphological categories are cell lines divided into?

A
  1. fibroblastic
  2. epithelial like cells
  3. lymphoblast like cells
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8
Q

What are fibroblastic cells?

A
  1. mammalian cell line
  2. multipolar/bipolar
  3. elongated shape that grows onto substrate
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9
Q

What are epithelial-like cells?

A
  1. mammalian cell line
  2. polygonal shape
  3. attach to substrate in discrete patches
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10
Q

What are lymphoblast like cells?

A
  1. mammalian cell line
  2. spherical shape
  3. grow in suspension, not surface attachment
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11
Q

What are the advantages of cell cultures?

A
  1. observe cell behavior without animal variations
  2. maintain cells for generation= reproducibility
  3. controlled growth environment
  4. exposure to specific isolated reagents
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12
Q

What are the disadvantages to cell culture growth?

A
  1. require standardized technique
  2. time consuming
  3. limited material
  4. loss of original cellular mechanisms
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13
Q

Where can cell cultures be applied to life and sciences?

A
  1. cell/gene function research
  2. biological product formation
  3. testing
  4. regenerative medicine
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14
Q

Why is protein purification important?

A
  1. study structure and function of specific protein
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15
Q

What is recombinant DNA technology used for?

A

overexpress a specific protein making it easier to purify

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16
Q

What are the steps of sub-cellular fractionation?

A
  1. mechanically blend tissue, to form homogenous mix
  2. centrifuge
  3. cell lysis
  4. ultracentrifugation
17
Q

What techniques can be used to lyse cells?

A
  1. osmotic shock
  2. ultrasonic vibration
  3. mechanical blending
18
Q

What is ultracentrifugation used for?

A
  1. separate organelles
19
Q

What are lipid rafts?

A
  1. PM domains high in cholesterol, sphingolipids, gangliosides
  2. detergent insoluble
  3. center for signal transduction
  4. abnormal protein processing in neurodegenerative disorders
20
Q

What are the different matrices used for column chromatography?

A
  1. ion-exchange
  2. gel-filtration
  3. affinity
21
Q

What is ion-exchange chromatography?

A

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

22
Q

What is gel-filtration chromatography?

A

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
Q

What is affinity chromatography?

A

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
Q

How are genetically engineered proteins formed?

A
  1. use of recombinant DNA technology

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

25
Q

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

A
  1. unfolds the proteins
  2. attaches uniform negative charge
  3. contains beta-mercaptoethanol to reduce disulfide bonds
26
Q

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

A
  1. the size of the denatured protein
27
Q

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

A
  1. isoelectric points and the pH of the solution
28
Q

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

A

the protein will migrate towards the + pole

29
Q

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

A

the protein will migrate towards the - pole

30
Q

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

A

it will not move.

31
Q

What is Sirt3 and what happens if it malfunctions?

A
  1. mitochondrial protein deacetylase

2. malfunciton= increased acetylated Sirt3 substrates

32
Q

What is Western Blotting?

A
  1. type of protein analysis that uses Ab-Ag interaction in order to determine a protein.
33
Q

What is indirect immuno-cytochemistry?

A
  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
Q

How is ELISA useful in protein analysis?

A
  1. provides protein detection and quantification
35
Q

How is ELISA used to determine presence of Ag?

A
  1. Ab-enzyme complex binds with the Ag, forming a color and absorbance level is then analyzed with machine
36
Q

When would Indirect ELISA be used?

A
  1. used to detect the presence of an Ab
  2. used for HIV infection
  3. use enzyme linked Ab react with Ag
37
Q

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

A
  1. enzyme-linked antibodies were bound to human antibodies and implies the Pt has antibodies to viral antigen
38
Q

What is sandwich ELISA?

A
  1. detects the antigen and the quantity of Ag.

2. uses standard curve in order to determine the amount of Ag present

Molecular BIO (24 decks)

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