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MBD > Techniques in protein analysis > Flashcards

Techniques in protein analysis Flashcards

1
Q

Includes the analysis, categorization of overall protein signatures of a genome

A

Proteomics

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2
Q

3 goals of Proteomics

A

Identify/compare proteins
Study the interactions of proteins
Predict cell behavior / develop drug target

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3
Q

This method is based on charge

A

Ion exchange chromatography (IEC)

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4
Q

This method is based on size

A

Size exclusion chromatography

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5
Q

Based on specific binding interaction

A

Affinity chromatography

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6
Q

Based on highly specific antibody-antigen interaction

A

ELISA

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7
Q

Western blot is done by ____, ____, and ____

A

Separation
Binding
Staining

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8
Q

SDS-PAGE separates proteins based on ______

A

Molecular weight

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9
Q

SDS-PAGE is also known as

A

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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10
Q

It is a classical method based on molecular weight and charge

A

Two-dimensional gel electrophoresis (2-DE)

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11
Q

A modified 2-DE by fluorescent dye binding

A

Two-dimensional differential gel electrophoresis (2D-DIGE)

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12
Q

Why do smaller proteins migrate faster?

A

Less resistance to gel matrix

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13
Q

A detergent with a strong protein denaturing effect

A

Sodium dodecyl sulfate (SDS)

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14
Q

It forms a mesh-like matrix

A

Polyacrylamide

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15
Q

(True or False)

pH below their pI carry a net negative charge

A

(False)
Below pI = positive
Above pI = negative

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16
Q

pH at which the net charge of the protein is zero

A

Isoelectric point

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17
Q

pI of the protein is also called

A

Isoelectric point

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18
Q

Proteins are adsorbed into wells by Immobilization thru ____ & _____

A

Hydrophobic interaction

Covalent bonds

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19
Q

Various antigen-antibody combinations are used in this method

A

ELISA

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20
Q

The enzyme activity in ELISA is measured ______

A

Colorimetrically

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21
Q

4 types of ELISA

A

Direct
Indirect
Sandwich
Competitive

22
Q

It is also known as the simplest ELISA

23
Q

The color change in direct ELISA using a spectrophotometer

24
Q

How many antibodies are involved in direct ELISA

25
Q

The most popular ELISA

26
Q

It uses an extra incubation step

A

Indirect ELISA

27
Q

It is designed for detection of soluble antigens

A

Sandwich ELISA

28
Q

(True or False)

In Indirect sandwich ELISA , the antigen is labelled with a second labelled antibody

A

False

labelled with unlabelled antibody

29
Q

Used when antigens are contaminated with proteins

A

Sandwich ELISA

30
Q

Useful when the target is a small molecule

A

Competitive ELISA

31
Q

Rapid, accurate, cost-effective method of microbial identification

A

MALDI-TOF MS

32
Q

The mass analyzer in MALDI-TOF MS

A

Time-of-flight (TOF)

33
Q

Ion source in MALDI-TOF MS

34
Q

This laser is rarely used

A

Transversely Excited Atmospheric (TEA-CO2)

35
Q

Nd:Yag stands for

A

Neodymium-doped yttrium aluminum garnet

36
Q

Er:Yag stands for

A

Erbium-doped yttrium aluminum garnet

37
Q

Matrix is used to ___ & ___ analyte molecules

A

dilute & isolate

38
Q

Ions of different m/z (mass-to-charge) are dispersed in a time during their flight of known length

A

Time-of-flight

39
Q

Consists of a series of ring electrodes with high voltage

A

Reflectron

40
Q

(True or False)

Faster ions carrying more kinetic energy will travel a shorter path

A

False

Longer path

41
Q

This analyzer is not suitable for analytes that are not stable enough to survive the electric field

A

Reflectron TOF analyzer

42
Q

It leads to ionization and sublimation of both sample and matrix

43
Q

Basic and important for protein characterization

A

Intact Mass determination

44
Q

It has widespread uses in proteomics to identify proteins from simple mixtures

A

Peptide Mass Fingerprinting (PMF)

45
Q

It is generated by digesting proteins of interest with a sequence specific enzyme

46
Q

It is used to make wells or lanes to load samples

47
Q

What do you use to clean glass plates before assembling the gel casting mold

48
Q

This should be avoided when pouring acrylamide solution because it inhibits polymerazation

A

Contact w/ air (oxygen)

49
Q

Allow acrylamide to polymerize for ___ minutes

50
Q

It is highly concentrated when migrating thru a stacking gel prior to entering a separating gel

MBD (3 decks)

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