FINALS - Protein Analysis

Question 1

what is the most abundant macromolecules in the biological system

Answer 1

proteins

Question 2

proteins are polymers comprising amino acid that are linked by ______

Answer 2

peptide bonds

Question 3

True or False:

proteins serve as:
1. antibodies
2. enzymes
3. messengers
4. structural components
5. transport
6. transport or storage molecule

Answer 3

true

Question 3

the proteins that can be detected in the blood of the patient are called?

Answer 3

biomarkers

Question 4

these are measurable substance in the blood whose presence is indicative of diseases or environmental exposure

Answer 4

biomarkers

Question 5

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

Answer 5

column chromatography

Question 6

this 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

what does the column chromatography uses to separate proteins based on their physical properties?

Answer 7

glass or plastic tube with resin inside

Question 8

what should be added in the column for proteins to migrate?

Answer 8

buffer solution

Question 9

the proteins the sample of a column migrate depending on?

Answer 9

1. nature of the matrix
2. physical properties
3. chemical properties

Question 10

in this protein analysis technique, the resin bead has many tiny pores like a WHIFFLE BALL

Answer 10

gel permeation/filtration/size exclusion

Question 11

in gel permeation, the molecules are separated based on their difference in?

Answer 11

size and shape

Question 12

in gel permeation, small molecules are more likely to go through the?

Answer 12

pore of the matrix

Question 13

true or false:

in gel filtration, large molecules move through the column more slowly since they cannot fit into the beads

Answer 13

false; quickly

Question 14

separates proteins using resin according to their SURFACE CHARGES

Answer 14

ion exchange chromatography

Question 15

it uses the LOCK AND KEY BINDING that is widely present in biological system

Answer 15

affinity chromatography

Question 16

it is used to separate and prepare LARGER quantities of PROTEINS and ANTIBODIES

Answer 16

affinity chromatography

Question 17

it uses the principle that the protein binds to molecules for which it has SPECIFIC AFFINITY

Answer 17

affinity chromatography

Question 17

this is the specific affinity where the PROTEIN BINDS TO CARRY OUT BIOLOGICAL ACTIVITY

Answer 17

ligand

Question 18

its purpose are for SEPARATION, to determine SIZES, PRESENCE/AMOUNT OF DNA

Answer 18

electrophoresis

Question 19

in electrophoresis, what is used to carry out proteins with a STACKING GEL and SEPARATING GEL?

Answer 19

polyacrylamide gel electrophoresis (PAGE) with SDS

Question 20

this technique is associated with STAINING METHOD and can DETECT BAND OF PROTEIN in a simple and relatively rapid manner

Answer 20

electrophoresis

Question 21

this DISRUPTS the structure of protein to be LINEAR and bind most protein

Answer 21

sodium dodecyl sulphate (SDS)

Question 22

give the agents that is used in SDS for denaturation and breaking of covalent bonds

Answer 22

dithiothreitol (DTT) or 2-mercaptoethanol

Question 23

this is often used to determine the MOLECULAR WEIGHT of proteins

Answer 23

SDS

Question 24

each sodium dodecyl sulfate contributes to how many negative charges?

Answer 24

two

Question 25

can detect as low as 10 ng - 1 ng of protein bands

Answer 25

colorimetric

Question 26

organic dye and most frequently employed method for protein detection in SDS-PAGE gel

Answer 26

coomassie blue staining

Question 27

uses IMIDAZOLE AND ZINC SALTS for protein detection in electrophoresis gels

Answer 27

zinc-reverse staining/negative staining

Question 28

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

spots in silver staining are _____ on a ____ background

Answer 29

dark brown to black; light beige

Question 30

what is used to solve differentially sensitive proteins to silver staining?

Answer 30

glutaraldehyde free modified silver staining

Question 31

it bands FLUORESCE

Answer 31

fluorescence

Question 31

better resolving power as compared to the SDS-PAGE

Answer 31

two-dimensional gel electrophoresis

Question 32

the most popular and well-established technique for GLOBAL PROTEIN SEPARATION AND QUANTIFICATION

Answer 32

gel-based proteomics

Question 33

this is an example of a gel-based proteomics

Answer 33

two-dimensional gel electrophoresis

Question 34

protein in 2DE is electrophoresed through a narrow tube gel containining molecules called?

Answer 34

ampholytes

Question 35

ampholytes set up a _____

Answer 35

pH gradient

Question 36

this is the pH at which protein has no net charge/neutral

Answer 36

isoelectric point

Question 37

results after isoelectric focusing and SDS-PAGE are visualized and quantified through?

Answer 37

bioinformatic software

Question 38

northern blot is for?

Answer 38

RNA

Question 39

southern blot is for?

Answer 39

DNA

Question 40

western blot is for?

Answer 40

proteins

Question 41

this is a laboratory method used to detect SPECIFIC PROTEIN molecules among a MIXTURE of proteins

Answer 41

western blot

Question 42

what solid support is used in western blotting in transferring the gel

Answer 42

nitrocellulose/nylonsheet/or PVDF (polyvinyl vinylidene difluoride)

Question 43

this mode of transferring proteins is aided by capillary action or vacuum

Answer 43

diffusion/passive transfer

Question 44

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

Answer 44

diffusion/passive transfer

Question 45

mode of transferring proteins by putting a gel membrane in a suitable holder and immersing both the gel and membrane in a tak filled with buffer fitted with TWO PLATE ELECTRODE (- and +)

Answer 45

electroblotting/electrotransfer

Question 46

these binds to the non-specific protein binding sites to prevent the binding of the antibodies to these sites

Answer 46

blocking agents

Question 47

these generates a colored product that could be conjugated to an enzyme

Answer 47

horseradish peroxidase (HRP) and alkaline phosphatase

Question 48

almost same with the color formation wherein it provides HIGHER SENSITIVITY than the COLOR PRODUCTION system

Answer 48

chemiluminescent

Question 49

in UV absorption method, what can be used in detecting proteins?

Answer 49

spectrophotometer

Question 50

the principle of this method is that nucleic acids (DNA or RNA) contain conjugated double bonds in their purine and pyrimidine rings that have a specific ABSORPTION PEAK at 260 nm

Answer 50

UV absorption peak

Question 51

the most commonly used reference protein in colorimetric

Answer 51

bovine serum albumine (BSA)

Question 52

in this method the peptide bonds of protein react with CUPRIC IONS present in BIURET reagents under alkaline condition

Answer 52

biuret method

Question 53

if cupric ions reacvct with biuret reagents it will produce a _____ color

Answer 53

purple

Question 54

presence of protein in biuret method will produce what color

Answer 54

violet

Question 55

this is based on the BIURET REACTION with ADDITIONAL steps and reagents to increase the sensitivity of detection

Answer 55

lowry protein assay

Question 56

in the biuret reaction, copper interacts with four nitrogen atoms of peptides that forms ____

Answer 56

cuprous complex

Question 57

reacts with cupric ions generated by the BIURET REACTION under ALKALINE condition forming BCA CUPROUS COMPLEX forming a purple color

Answer 57

bicinchonic acid method

Question 58

this is a common laboratory technique which is used to MEASURE THE CONCENTRATION of an ANALYTE in a solution

Answer 58

ELISA

Question 59

ability to DISCRIMINATE among DIVERSE protein

Answer 59

specificity

Question 60

ability to TIGHTLY BIND to molecules

Answer 60

afiinity

Question 61

what produces the chemical interaction between antibodies in ELISA

Answer 61

B cells of leukocytes and antigens

Question 62

y-shaped; any foreign substances

Answer 62

antigen

Question 63

green color attached to antigen

Answer 63

antibody

Question 64

uses a primary labeled antibody that react DIRETCLY with the antigen

Answer 64

direct ELISA

Question 65

it only uses an enzyme-labelled PRIMARY antibody; SECONDARY antibodies are not needed

Answer 65

direct ELISA

Question 66

it utilizes a primary UNLABELED antibody in conjunction with a labeled SECONDARY antibody

Answer 66

indirect ELISA

Question 67

antigens like TUMOR MARKERS, HORMONES, SERUM PROTEINS may be determined and bind with the capture antubody and become IMMOBILIZED

Answer 67

sandwich ELISA

Question 68

uses to determine the PRECISE MASS of PEPTIDES

Answer 68

MALDI-TOF spectrometry

Question 69

The peptides are immobilized in an organic MATRIX and then blasted with a LASER, causing them to be ejected in the form of an ionized gas.

Answer 69

MALDI-TOF