22) Unit 1 - Antibody Techniques & Microscopy

Question 1

what is the purpose of antibody techniques

Answer 1

using antibodies doe the detection and identification of proteins

Question 2

Name the two types of antibody techniques

Answer 2

• immunoassay

* protein blotting

Question 3

what is immunoassay

Answer 3

Branch of biology that focuses on the use of antibodies to detect the presence and concentration of a protein in a solution

Question 4

How does immunoassay work?

Answer 4

• The antibody will be attached to a detectable label which shows when binding has occurred (often colour changing enzyme)
• When binding of the antibody to the antigen then there will be a colour change

Question 5

uses for immunoassay and protein blotting?

Answer 5

• detection of HIV

* testing for drugs (not protein blotting just immuonassay)

Question 6

Why is protein blotting used??

Answer 6

allows proteins from a tissues or cell to be extracted and detected

Question 7

Process of protein blotting?

Answer 7

• Protein sample (tissues/cells) are first separated using gel electrophoresis
• Proteins are then transferred to a membrane
• Membrane probed for protein of interest using specific antibody, linked to reporter enzyme (e.g.florescence)

Question 8

what is the meaning of immunohistochemistry

Answer 8

when antibodies are used to detect the presence of a particular antigen within a tissue sample

Question 9

why would you use monoclonal antibodies and how are they made

Answer 9

why?:
•To produce a large quantity of a particular antibody

How?:
Hybridomas are formed from the fusion of a B lymphocyte and myeloma cell using polyethylene glycol (PEG)

Question 10

uses of monoclonal antibodies?

Answer 10

Diagnosis and detection of a disease? e.g HIV

Question 11

what is bright field microscopy

Answer 11

When light is transmitted through a specimen or sample to an objective lens which magnifies the image and which is then shown in the eyepeice

Question 12

use of bright field microscopy?

Answer 12

• examine whole organisms
• examine parts of an organism
• examine thing sections of dissected tissue

Question 13

use of fluorescence microscopy

Answer 13

allows particular protein structures to be visualized

Question 14

why use haemocytometry

Answer 14

to estimate the concentration of cells in a sample using a cell count slide

Question 15

why use flow cytometry

Answer 15

allows cells to be detected, counted and analysed one by one