TECHNIQUES IN PROTEIN ANALYSIS

Question 1

it is the most abundant macromolecules in biological system.

Answer 1

Proteins

Question 2

They are polymers comprising amino acids that are linked by peptide bonds.

Answer 2

Proteins

Question 3

These proteins can be detected in the blood of the patient which are considered as __________

Answer 3

Biomarkers

Question 4

it is a measurable substance in the blood whose presence is indicative of diseases or environmental exposure

Answer 4

biomarkers

Question 5

it is the most common method for purifying proteins from other protein molecules with a given sample

Answer 5

Column Chromatography

Question 6

it involves the separation of soluble components in a solution by specific differences in physical-chemical characteristics of the different constituents

Answer 6

Column Chromatography

Question 7

True or False:
Small molecules are more likely to go through the pore of the matrix. They are trapped inside the resin and will travel through the column more slowly.

Answer 7

True

Question 8

True or False:
Large molecules move through the column more quickly

Answer 8

True

Question 9

This type of column chromatography has many tiny pores in the resin bead.

Answer 9

Gel/ Gel Permeation/ Gel Filtration/ Size Exclusion

Question 10

This type of column chromatography uses resin to separate proteins according to their surface charges

Answer 10

Ion Exchange Chromatography

Question 11

It contains a resin bearing either positively or negatively charged chemical groups.

Answer 11

Ion Exchange Chromatography

Question 12

True or False:
In Ion Exchange Chromatography, the resins with positively charged groups (anion-exchange resins) attract negatively charged solute.

Answer 12

True

Question 13

True or False:
In Ion Exchange Chromatography, Opposite charges attract. - It traps (-) charged molecules so (+) charged
molecules are first obtained.

Answer 13

True

Question 14

This type of chromatography uses the so called lock and key binding that is widely present in biological system.

Answer 14

Affinity Chromatography

Question 14

It is used to separate and prepare larger quantities of proteins and antibodies for study.

Answer 14

Affinity Chromatography

Question 14

This type of Column Chromatography It uses the principle that the protein binds to a
molecules for which it has specific affinity.

Answer 14

Affinity Chromatography

Question 15

This technique has different purposes: : for separation, to determine the sizes, presence, or amount of DNA

Answer 15

Electrophoresis

Question 16

We use this kind of gel electrophoresis for proteins

Answer 16

Polyacrylamide gel electrophoresis with SDS

Question 17

It is carried out with a stacking gel and separating gel

Answer 17

Polyacrylamide gel electrophoresis with SDS

Question 18

This technique associated with staining method can detect bands of protein in a simple and relatively rapid manner.

Answer 18

Electrophoresis

Question 19

SDS-PAGE meaning

Answer 19

Sodium Dodecyl Sulphate Polyacrylamide gel electrophoresis

Question 20

True or False:
Sodium Dodecyl Sulphate Polyacrylamide gel electrophoresis is an anionic detergent (-)

Answer 20

True

Question 21

It disrupts the structure of protein to be linear and binds most protein

Answer 21

Sodium Dodecyl Sulphate

Question 22

These agents break the covalent bonds that are present

Answer 22

dithiothreitol (DTT) or 2-
mercaptoethanol

Question 23

Advantages of SDS

Answer 23

it coats all the polypeptides with negative charges It masks the natural charges of the subunit

Question 24

What are the two types of staining methods

Answer 24

Colorimetric and Fluorescence

Question 25

Give the 3 high sensitivity colorimetric staining methods that can be used either directly after electrophoresis

Answer 25

Coomassie blue staining, zinc-reverse staining, and silver staining

Question 26

It is one of the 3 high sensitivity colorimetric that is most frequently employed method for protein detection in SDS-PAGE gel

Answer 26

Coomassie blue staining

Question 27

It is one of the 3 high sensitivity colorimetric that Uses imidazole and zinc salts for protein detection in electrophoresis gels

Answer 27

Zinc-reverse staining/ Negative Staining

Question 28

It is one of the 3 high sensitivity colorimetric that is based on the binding of silver ions to the proteins followed by reduction to free silver, sensitization, and enhancement

Answer 28

Silver staining

Question 29

This dye give very wide linear dynamic ranges, over four orders of magnitude.

Answer 29

Fluorescence dye

Question 30

Although this type of dye can even reveal as low as 1 ng of protein, it is inconvenience because it has to be incorporated before gel electrophoresis.

Answer 30

CyeDyes

Question 31

It is a better resolving power as compared to the SDSPAGE, especially if there is a mixture of polypeptides or when it’s too complex

Answer 31

Two-dimensional Gel Electrophoresis

Question 32

the complex protein samples are separated in two dimensions according to their net charge at different pH and their molecular weights determined by SDS-PAGE.

Answer 32

Two-dimensional Gel electrophoresis

Question 33

In this dimension, the . Sample is placed in a narrow tube gel or subjected to isoelectric focusing based on pH.

Answer 33

First Dimension

Question 34

In this dimension, the strip is placed at the top of the SDS-PAGE.

Answer 34

Second dimension

Question 35

It is another method related to gel electrophoresis that is used to detect specific protein molecules from among a mixture of proteins

Answer 35

Western Blot

Question 36

This blot is used to detects the RNA

Answer 36

Northern blot

Question 37

This blot is used to detects the DNA

Answer 37

Southern blot

Question 38

This blot is used to detects the Proteins

Answer 38

Western blot

Question 39

3 elements used in western blot

Answer 39

1. Separation by size. 2. Transfer to a solid support. 3. Use a specific or proper primary and secondary antibody for visualization.

Question 39

It is a method of transferring protein onto membranes that is aided by capillary action (by placing a weight on the gel and membrane) or vacuum (for faster and more uniform transfer)

Answer 39

diffusion/ passive transfer

Question 40

It is achieved by posing a membrane sheet on one or both side of the gel

Answer 40

diffusion/ passive transfer

Question 41

This method of transferring protein onto membranes are driven by an electrical field and is achieved by putting a gel membrane in a suitable holder and immersing both the gel and membrane in a tank filled with buffer fitted with two plate electrode (+ and -)

Answer 41

electroblotting/ electrotransfer

Question 42

This method of transferring protein can use spectrophotometer in detecting proteins

Answer 42

UV Absorption Method

Question 43

A common laboratory technique which is used to measure the concentration of an analyte in a solution

Answer 43

ELISA

Question 44

It is a quantitative immunological procedure in which antigen-antibody reaction is monitored by enzyme measurement.

Answer 44

ELISA

Question 45

This assay uses the coupling of antigens and antibodies and relies on the specificity and affinity of antibodies for antigens.

Answer 45

ELISA

Question 46

It uses a primary labeled antibody that react directly with the antigen

Answer 46

DIRECT ELISA

Question 47

It can be performed with the antigen that is directly immobilized on assay plate.

Answer 47

DIRECT ELISA

Question 48

it uses only an enzyme-labelled primary antibody. The secondary antibodies are not needed.

Answer 48

DIRECT ELISA

Question 49

It utilizes a primary unlabeled antibody in conjunction with a labeled secondary antibody

Answer 49

INDIRECT ELISA

Question 50

both primary and secondary antibody are used, but in this case, the primary antibody is not labelled (no enzyme-linked). The enzyme is labelled with the secondary antibody.

Answer 50

INDIRECT ELISA

Question 51

Antigens like Tumor markers, hormones, serum proteins may be determined.

Answer 51

SANDWICH ELISA

Question 52

In this type of ELISA, you need to prepare a surface to which a known quantity of antibody is bound.

Answer 52

SANDWICH ELISA

Question 53

it is Used to determine the precise mass of peptides.

Answer 53

MALDI-TOF

Question 54

IT is a very sensitive and accurate for determining amino acid sequences.

Answer 54

MALDI-TOF