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1.1 Introduction To Biomedical Science > Antibodies - Uses > Flashcards

Antibodies - Uses Flashcards

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Biology 101 flashcards
1
Q

List the uses of antibodies

A

Immunostaining

Flow Cytometry

Immunoassays

Immunoprecipitation

Western blotting

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2
Q

Why is immunostaining done?

3

A

To detect cell and tumour proteins

To identify micro-organisms - detect the presence of some diseases

To investigate cell phenotype

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3
Q

How is immunostaining carried out?

A

Antibody labelled with fluorescent dye or enzyme which creates coloured deposit at site of antigen

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4
Q

In what lab is immunostaining carried out?

A

Histology laboratory

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5
Q

How can immunostaining be used for cell and tumour identification?

A

It can tell us where the primary tumour is from

It can tell us what tumour subtype it is

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6
Q

Explain in your own words what immunostaining is.

3

A

An antibody is used to stain a sample

The antibody is labelled with fluorescent dye or an enzyme

A coloured deposit is created at the site of the antigen - a specific protein epitope

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7
Q

What is flow cytometry

A

A method of cell sorting and quantitation

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8
Q

Explain how flow cytometry works in your own words.

A

Similar to water going down a sink

As it spins the cells are in a line that passes by a laser

A flash will occur when the selected antibody and attached cell pass by the laser

The number of the cell type is counted as each one passes by the laser

The machine will also separate the chosen cell type from the sample

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9
Q

How is the cell separated from the sample in flow cytometry?

2

A

As the chosen cell passes through the device a charge is applied to the cell

The machine then pulls the chosen cell away from the sample

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10
Q

Give an example of a type of cell used in flow cytometry?

A

Stem cells

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11
Q

What antibody is used to count stem cells?

A

CD-34

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12
Q

What is an immunoassay also called?

A

Ligand-binding assay

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13
Q

What is the basis of immunoassays?

A

A broad range of techniques whose common feature is that measurement is based on specific recognition of the analyte by an antibody or other macromolecule

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14
Q

What are the four main immunoassay methods?

A

Radio-labelled RIA

Enzyme-labelled EIA

Chemiluminescence

Fluorescence- labelled

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15
Q

What is a radio-labelled RIA?

2

A

An antibody is labelled with radioactive isotope

The radioactivity is measured

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16
Q

Why is a radio-labelled RIA rarely used?

A

It is dangerous and needs a special lab

17
Q

What is an enzyme-labelled EIA?

3

A

ELISA

An antibody is labelled with an enzyme

Colour reaction is measured spectrophotometrically

18
Q

Why is an ELISA not often used?

A

Enzymes are too valuable to be used

19
Q

What is chemiluminescence?

A

An antibody is labelled with chemiluminescent dye

20
Q

What is fluorescence labelled?

2

A

An antibody is labelled with fluorescent dye

The fluorescence is measured

21
Q

What are immunoassays used for?

2

A

Measuring blood/fluid constituents

Detection of specific proteins in fluid samples

22
Q

What is immunoprecipitation used for?

A

The detection of proteins and bacteria

23
Q

How is immunoprecipitation used to detect bacteria?

A

Bacteria are detected using antigen-antibody reactions

24
Q

Explain in your own words how immunoprecipitation works - using Syphilis as an example
(6)

A

Latex beads are coated in anti syphilis antibody

Syphilis reacts with beads

This causes beads to clump together

Drop of latex beads and drop of patients plasma

Drops will disperse and stain the gel

If a line appears between antigen and antibody then the test is positive

25
Q

What are the two different ways of carrying out immunoprecipitation?
(2)

A

Using one drop of the latex beads coated in anti syphilis antibody and one drop of the patients plasma

One drop with antigen and one drop with antibody

26
Q

Explain in your own words how western-blotting works.

6

A

Electrophoresis - plastic is put on gel

The gel is dried out

Bands stick to plastic

Stain with antibody which has a radiolabel in it

Photograph it

Antigen bands develop on film

1.1 Introduction To Biomedical Science (11 decks)

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