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BIOC 3622 - Biochemistry Lab > Gel Filtration Chromatography > Flashcards

Gel Filtration Chromatography Flashcards

1
Q

______________ is a very powerful technique for the physical separation of molecules on the basis of size.

A

Size exclusion chromatography

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2
Q

What is a molecular disease of hemoglobin?

A

Sickle cell anemia

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3
Q

Hemoglobin is made up of ______________ which is associated to form a large, globular protein.

A

4 polypeptides (small proteins)

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4
Q

Hemoglobin has a color _____ and has a molecular weight of ________.

A

brown; 65000 daltons

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5
Q

The mass of beads within the column is often referred to as the _________.

A

column bed

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6
Q

This kit contains 8 columns which are prefilled with beads that effectively ___________ molecules that are below 60,000 daltons.

A

separate (“fractionate”)

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7
Q

__________ allows the separation of individual components from complex mixtures.

A

Chromatography

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8
Q

What is used in the stationary phase for paper chromatography?

A

paper

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9
Q

What is the mobile phase?

A

the solvent and the molecules to be separated

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10
Q

In step 1 of the procedure, the last two tubes will be labeled as _________ and __________.

A

Waste; Column Buffer

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11
Q

The liquid used to dissolve the biomolecules to make a mobile phase is called a _____.

A

buffer

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12
Q

Hemoglobin has a higher affinity for _____ than oxygen.

A

carbon monoxide

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13
Q

What kind of chromatography separates molecules based on charge?

A

Ion-exchange chromatography

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14
Q

Vitamin B12 is _____ and has a molecular weight of _______.

A

pink; 1350 daltons

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15
Q

At high altitudes, where the amount of oxygen is decreased, the body responds by increasing the number of _______ produced.

A

red blood cells

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16
Q

Larger molecules are ____ from the beads.

A

“excluded”

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17
Q

How is chromatography commonly used in biotechnology?

A

* purifying biological molecules like proteins;

* for medicine;

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18
Q

In ____________, a biomolecule (often an antibody) will bind to the protein to be purified and is attached to the beads.

A

affinity chromatography

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19
Q

In simple terms, what does chromatography accomplish?

A

It allows the separation of individual components from complex mixtures.

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20
Q

In step 2, how many ml of column buffer should be pipetted into the tube labeled column buffer?

A

4 mL

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21
Q

In ion exchange chromatography, if the beads is positive, it will bind negatively charged ions. This technique is called _________.

A

anion exchange

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22
Q

In step 8, when all of the liquid has been drained from the column, what should be added to the top of the column?

A

3ml of column buffer

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23
Q

_______. a protein found in red blood cells, functions to transport ______ to the tissues of the body.

A

Hemoglobin; oxygen

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24
Q

________ and ___________ are the two biomolecules in your sample.

A

Hemoglobin; vitamin B12

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25
Q

The mixture of biomolecules dissolved in the buffer is called a

A

sample

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26
Q

Microscopic porous spheres

A

beads

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27
Q

What happens in Fraction 3?

A

The larger protein molecules emerge from the column first.

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28
Q

In affinity chromatography, the elution buffer often contains ________.

A

high concentrations of salt or acid

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29
Q

Sources rich in vitamin B12 include

A

eggs, dairy products and meats

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30
Q

When the contaminant is separated from the protein of interest, a ____________ is used to get the desired protein to elute from the column.

A

high salt buffer

31
Q

In step 7, when all the liquid has been drained from the column, what should be added?

A

Another 250 ul of column buffer

32
Q

The side effects of sickle cell anemia can be alleviated by ____________ from people who have normal hemoglobin and red blood cells.

A

frequent blood transfusions

33
Q

The beads act as ______ or _____ and function to filter small molecules which become temporarily trapped within the pores.

A

“traps”; “sieves”

34
Q

The hemoglobin used in this experiment is _______.

A

bovin hemoglobin

35
Q

What is used in the stationary phase for column chromatography?

A

glass beads called resin

36
Q

The larger molecules which pass quickly through the column will end up in the early tubes or _____.

A

“fractions”

37
Q

_________ is a vitamin essential to humans and other vertebrates.

A

Vitamin B12

38
Q

True or false? Vitamin B12 is not found in plants and vegetable foods.

A

True

39
Q

Hemoglobin can bind to ______ and _______.

A

oxygen; carbon monoxide

40
Q

Chromatography consists of a _____ phase and a ______ phase.

A

mobile; stationary

41
Q

In step 4, why should the column not be jammed tightly into the collection tubes?

A

The column will not flow

42
Q

Molecules travel through the stationary phase at different rates because of their ______.

A

chemistry

43
Q

What is the main component of red blood cells?

A

Hemoglobin

44
Q

Gel filtration chromatography is commonly referred to as…

A

size exclusion chromatography.

45
Q

A mixture of large and small ____ is applied to a column of porous beads.

A

proteins

46
Q

How does the Vitamin B12 complex pass through the intestine and into the blood stream?

A

It binds to the carrier protein

47
Q

As the buffer flows down the column, the small _____ molecules penetrate into the ____ and are ____.

A

protein; beads; slowed

48
Q

The ___________ emerge from the column first.

A

larger protein molecules

49
Q

In ion exchange chromatography, when the contaminant is separated from the protein of interest, a _______ is used to get the desired protein to elute from the column.

A

high salt buffer

50
Q

in Fraction 1, what is happening?

A

A mixture of large and small proteins are applied to a column of porous beads.

51
Q

In step 6, after adding the protein mix, what should be added next to the top of the column?

A

250 ul of column buffer

52
Q

One function of vitamin B12 is ______________.

A

the breakdown of fats

53
Q

In step 9, when _____ have been collected into tube 2, transfer the column onto ____.

A

5 drops of buffer; tube 3

54
Q

Molecules greater than 60,000 pass around the beads and are excluded from the column — also referred as to as the _________ of a column.

A

exclusion limit

55
Q

Why are muscles that are very active and require a lot of oxygen dark in color?

A

High myoglobin content

56
Q

What is responsible for the red-brown color of hemoglobin?

A

iron-containing heme group

57
Q

As the liquid flows through the column, molecules below 60,000 daltons enter the beads and pass through the column more _____.

A

slowly

58
Q

In step 5, what should be dropped onto the column bed?

A

one drop of protein mix

59
Q

What kind of chromatography separates molecules based on size?

A

-Gel filtration/Size exclusion chromatography -Molecular sieve chromatography

60
Q

In this lab, what will be investigated?

A

Size exclusion chromatography.

61
Q

What happens in Fraction 2?

A

As the buffer flows down the column, the small protein molecules penetrate into the beads and are slowed.

62
Q

People who have a strict vegetarian diets are often deficient in what?

A

Vitamin B12.

63
Q

Once the fractions have been collected, they should be covered with ________.

A

parafilm

64
Q

In __________, if the beads are negatively charged, they bind to positively charged molecules.

A

cation exchange

65
Q

In ______________, microscopic beads which contain tiny holes are packed into a column.

A

gel filtration chromatography

66
Q

Vitamin B12 are required in large quantities. True or false?

A

No, they are only required in minute quantities

67
Q

In step 10, what should be done to the column when finishing collecting drops?

A

Cap it

68
Q

In affinity chromatography, what is the buffer used for?

A

To disrupt the bond between the protein of interest and the antibody

69
Q

In ion exchange chromatography, what is the purpose of the high salt buffer?

A

To get the desired protein to elute from the column

70
Q

As the liquid flows down the column, what happens to the molecules below 60000 daltons?

A

Molecules enter the beads and pass through the column more slowly.

71
Q

As the liquid flows down the column, what happens to the molecules above 60000 daltons?

A

Molecules pass around the beads are are excluded from the column.

72
Q

negatively charged resin; binds to positive

A

Cation exchanger

73
Q

positively charged resin; binds to negative

A

Anion exchanger

BIOC 3622 - Biochemistry Lab (8 decks)

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