Tissue Prep And Staining Contd

Question 1

What are immunocytochemical techniques?

Answer 1

Techniques used to study the presence of specific antigens by using monoclonal antibodies.

Question 2

Give examples of the antigens

Answer 2

Glycoproteins, proteins, and proteoglycans

Question 3

How are monoclonal antibodies made?

Answer 3

From B cells ( the B cells are very specific)

Question 4

What do antigens have many of, with respect to antibodies?

Answer 4

They have multiple epitopes (binding sites) that attract a number of different antibodies (polyclonal).

Question 5

How do you make a hybridoma and what is their advantage?

Answer 5

B cell + myeloma. They have the advantage of being very specific and the ability to grow indefinitely.

Question 6

What are the 3 ways of direct labeling of antibodies?

Answer 6

Fluorescent dye- visible in fluorescent microscope
Visible substance - visible in light microscope
Gold or ferritin- visible in electron microscope.

Question 7

What is indirect labeling of antibodies?

Answer 7

A marker is attached to a secondary antibody that is specific to the second antibody used to locate the antigen of interest.

Question 8

RNA rich organelles may be stained with which type of dye? And does a control have to be used?

Answer 8

Basic dyes and yes a control is used and incubated in ribonuclease