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AH biology - unit 1 (Proteomics) > Unit 1.1 - Lab techniques for Biologists > Flashcards

Unit 1.1 - Lab techniques for Biologists Flashcards

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1
Q

Give examples of COSHH

A

Chemicals
Products containing chemicals
Fumes
Dusts
Vapours
Biological agents

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2
Q

What does COSHH stand for?

A

Control of substances hazardous to health

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3
Q

What does PPE stand for

A

Personal protective equipment

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4
Q

Give examples of PPE

A

Safety glasses
Lab coat
Gloves

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5
Q

What does an autopipette allow to be measured

A

Small volumes of liquid accurately

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6
Q

What are burettes used for?

A

Dispense precise volumes of liquid reagants

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7
Q

What happens to burette before it is used

A

Rinsed with the liquid to be used

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8
Q

Where are readings taken from?

A

Bottom of meniscus

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9
Q

What is repeated dilution from a stock solution known as? (regular intervals)

A

Serial dilution

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10
Q

What type of dilution is it if each step is a tenfold dilution?

A

Log dilution

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11
Q

The concentration of a pigmented compound can be quantified using a?

A

Colorimeter

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12
Q

How does a colorimeter work?

A

Passing a light beam at a specific wavelength through a cuvette containing a sample solution

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13
Q

How do you calibrate a colorimeter?

A

Place a calibration solution(usually distilled water) into a cuvette, place in colorimeter and press calibrate, providing a user with a 0 absorbance reading

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14
Q

What is a pH buffer

A

Solution whose pH changes very little when a small amount of acid/base is added to it (to maintain pH)

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15
Q

Give one method to determine the unknown concentration of a solution

A

Standard curve (titration)

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16
Q

What seperates substances out in centrifugation?

A

Size and density

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17
Q

What is the liquid that remains above the pellet called in centrifugation?

A

Supernatant fluid

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18
Q

What does protein electrophoresis seperate proteins based on?

A

Charge/size

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19
Q

Give one form of protein electrophoresis

A

SDS page

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20
Q

Do smaller proteins travel furthur or less than bigger proteins in protein electrophoresis?

A

Further

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21
Q

What does seperation of proteins depend on if native gels are used?

A

Protein charge and shape

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22
Q

What do proteins do at their isoelectric point?

A

Overall neutral charge, precipitate out of solution

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23
Q

What charge do proteins carry below their isoelectric point?

A

Positive

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24
Q

How does isoelectric focusing work?

A

pH gradient is set up on gel:
Protein loaded in and moves until it reaches its isoelectric point where it forms a band

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25
Q

What is Chromatography?

A

Set of Techniques which separate the components of a mixture

26
Q

What are the three types of chromatography?

A

Paper, thin-layer & affinity
chromatography

27
Q

What is the stationary phase in paper chromatography?

A

Strip of chromatography paper

28
Q

What is the stationary phase in thin layer chromatography?

A

Strip of absorbent paper such as silica gel

29
Q

What does affinity chromatography rely on?

A

Binding interactions of protein and a ligand

30
Q

Where is the protein found in affinity chromatography?

A

Immobilised by specific receptor (ligand)

31
Q

What is immunoassay?

A

Test which uses antibodies to detect the presence and concentration of a protein within a solution

32
Q

What does immunoassay rely on

A

Specificity of antibodies

33
Q

What allows scientists to detect when binding has occurred in an immunoassay test?

A

Each antibody must be linked to a detectable label

34
Q

What does a reporter enzyme do?

A

Causes a colour change in the presence of a specific antigen

35
Q

What does ELISA stand for?

A

Enzyme linked immunosorbent assay

36
Q

What is an ELISA?

A

Analytical technique which uses antibodies to detect the presence of an antigen in a solution

37
Q

What are three types of ELISA?

A

Direct, Indirect, Sandwich

38
Q

What is the difference between direct and indirect ELISA?

A

Direct - Primary antibody binds to antigen and has a reporter protein on it
Indirect - Primary antibody binds to antigen then secondary antibody binds with a reporter enzyme

39
Q

What happens during Sandwich ELISA?

A

Capture antibody bound to surface of multiwell plate
Antigen is added and allowed to bind
Primary antibody added and binds
Secondary antibody, linked to a reporter enzyme added and binds to primary antibody

40
Q

Give examples of uses of ELISA

A

Detection of HIV
Detection of food allergies
Screening for certain types of drugs

41
Q

What does Protein Blotting allow?

A

Identify specific proteins seperated by gel electrophoresis

42
Q

What is the first step in protein blotting?

A

Proteins are separated using gel electrophoresis and transferred to a membrane

43
Q

How is the membrane probed for the protein of interest?

A

Using a specific antibody linked to a detectable label

44
Q

Give examples of applications of protein blotting

A

Identify specific proteins which are present in a cell sample
Detection of HIV and hepatitis B

45
Q

What does Fluorescence microscopy allow?

A

Particular protein structures to be visualised

46
Q

What is a fluorescent molecule?

A

One which absorbs a specific wavelength of light then emits a different (longer) wavelength

47
Q

How does Fluorescence microscopy work?

A

Specific proteins have fluorescent markers added to them
Cells then placed on a slide
Protein structures visualised using a fluorescence microscope

48
Q

What does a haemocytometer allow?

A

Estimation of the concentration of cells in a sample to be made

49
Q

What border can cells be touching to be counted?

A

Top and right border

50
Q

What can the reliability of a glass haemocytometer be effected by?

A

Incorrect placement of the cover slip

51
Q

Give a benefit of disposable haemocytometers?

A

Limit exposure to infectious materials

52
Q

Give examples of things a typical culture media contains

A

Water
Salts
Amino Acids
Vitamins
Glucose

53
Q

What do all animal cell cultures require?

A

Media containing growth factors

54
Q

Where do the growth factors come from?

A

Serum

55
Q

What is the inoculum?

A

Cells that have been used to inoculate culture media

56
Q

What are antibodies?

A

Y shaped proteins produced by B lymphocytes as part of the immune response

57
Q

What are antibodies used for?

A

Detect and Identify specific proteins

58
Q

What does fluorescent labelled protein blotting allow a scientist to identify?

A

Specific proteins that were seperated using gel electrophoresis

59
Q

What is fluorescent Immunohistochemical Staining?

A

Used to visualize distribution of specific cellular components in live cell

60
Q

What does separation rely on if non native gels are used in protein electrophoresis?

A

Size

61
Q

State two types of biological material that can be viewed by bright field microscopy

A

Thin sections of tissue
- Parts of an organism

62
Q

How are cells detached from tissues for cell lines?

A

Using proteolytic enzymes

AH biology - unit 1 (Proteomics) (11 decks)

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