Techniques in protein analysis

Question 1

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

Answer 1

Proteomics

Question 2

3 goals of Proteomics

Answer 2

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

Question 3

This method is based on charge

Answer 3

Ion exchange chromatography (IEC)

Question 4

This method is based on size

Answer 4

Size exclusion chromatography

Question 5

Based on specific binding interaction

Answer 5

Affinity chromatography

Question 6

Based on highly specific antibody-antigen interaction

Answer 6

ELISA

Question 7

Western blot is done by ____, ____, and ____

Answer 7

Separation
Binding
Staining

Question 8

SDS-PAGE separates proteins based on ______

Answer 8

Molecular weight

Question 9

SDS-PAGE is also known as

Answer 9

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Question 10

It is a classical method based on molecular weight and charge

Answer 10

Two-dimensional gel electrophoresis (2-DE)

Question 11

A modified 2-DE by fluorescent dye binding

Answer 11

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

Question 12

Why do smaller proteins migrate faster?

Answer 12

Less resistance to gel matrix

Question 13

A detergent with a strong protein denaturing effect

Answer 13

Sodium dodecyl sulfate (SDS)

Question 14

It forms a mesh-like matrix

Answer 14

Polyacrylamide

Question 15

(True or False)

pH below their pI carry a net negative charge

Answer 15

(False)
Below pI = positive
Above pI = negative

Question 16

pH at which the net charge of the protein is zero

Answer 16

Isoelectric point

Question 17

pI of the protein is also called

Answer 17

Isoelectric point

Question 18

Proteins are adsorbed into wells by Immobilization thru ____ & _____

Answer 18

Hydrophobic interaction

Covalent bonds

Question 19

Various antigen-antibody combinations are used in this method

Answer 19

ELISA

Question 20

The enzyme activity in ELISA is measured ______

Answer 20

Colorimetrically

Question 21

4 types of ELISA

Answer 21

Direct
Indirect
Sandwich
Competitive

Question 22

It is also known as the simplest ELISA

Answer 22

Direct

Question 23

The color change in direct ELISA using a spectrophotometer

Answer 23

Red

Question 24

How many antibodies are involved in direct ELISA

Answer 24

1

Question 25

The most popular ELISA

Answer 25

Indirect

Question 26

It uses an extra incubation step

Answer 26

Indirect ELISA

Question 27

It is designed for detection of soluble antigens

Answer 27

Sandwich ELISA

Question 28

(True or False) | In Indirect sandwich ELISA , the antigen is labelled with a second labelled antibody

Answer 28

False | labelled with unlabelled antibody

Question 29

Used when antigens are contaminated with proteins

Answer 29

Sandwich ELISA

Question 30

Useful when the target is a small molecule

Answer 30

Competitive ELISA

Question 31

Rapid, accurate, cost-effective method of microbial identification

Answer 31

MALDI-TOF MS

Question 32

The mass analyzer in MALDI-TOF MS

Answer 32

Time-of-flight (TOF)

Question 33

Ion source in MALDI-TOF MS

Answer 33

MALDI

Question 34

This laser is rarely used

Answer 34

Transversely Excited Atmospheric (TEA-CO2)

Question 35

Nd:Yag stands for

Answer 35

Neodymium-doped yttrium aluminum garnet

Question 36

Er:Yag stands for

Answer 36

Erbium-doped yttrium aluminum garnet

Question 37

Matrix is used to ___ & ___ analyte molecules

Answer 37

dilute & isolate

Question 38

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

Answer 38

Time-of-flight

Question 39

Consists of a series of ring electrodes with high voltage

Answer 39

Reflectron

Question 40

(True or False) | Faster ions carrying more kinetic energy will travel a shorter path

Answer 40

False | Longer path

Question 41

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

Answer 41

Reflectron TOF analyzer

Question 42

It leads to ionization and sublimation of both sample and matrix

Answer 42

MALDI

Question 43

Basic and important for protein characterization

Answer 43

Intact Mass determination

Question 44

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

Answer 44

Peptide Mass Fingerprinting (PMF)

Question 45

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

Answer 45

Peptides

Question 46

It is used to make wells or lanes to load samples

Answer 46

Comb

Question 47

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

Answer 47

Ethanol

Question 48

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

Answer 48

Contact w/ air (oxygen)

Question 49

Allow acrylamide to polymerize for ___ minutes

Answer 49

20-30

Question 50

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

Answer 50

Proteins