Immunocytochemistry

Question 1

What is immunocyto(histo)chemistry?

Answer 1

using Abs to detect/localize specific Ags in cells and/or tissues

Question 2

What is histology?

Answer 2

differential staining of cells in tissues with dyes (not Abs) to determine overall tissue morphology and the presence of cell types

Question 3

What is histochemistry?

Answer 3

cells are incubated with a substrate to indenitfy enzymatic activity in cells or to detect protein expression using enzyme-linked proteins (e.g. LacZ transgene)

Question 4

what is immunochemistry?

Answer 4

refers to the identification/quantification of Ags with specific Abs

Question 5

What is the purpose of dehydration and clearing during tissues fixation and emedding?

Answer 5

dehydration: removes fixative
clearing: accepts the embedder

Question 6

What is another word for eptiopes?

Answer 6

antigenic determinants

Question 7

What is a conformational determinant? What happens when the protein is denatured?

Answer 7

The Ab see the shape –> determinant lost by denaturation

Question 8

What is a linear determinant? What happens when the protein is denatured?

Answer 8

The Ab sees the sequence (already linearized) –> Ab still binds to its epitope when the protein is denatured (accessible determinant)

The Ab cannot see its sequence when the protein has shape –> Ab is able to bind to its epitope when the protein is denatured (inaccessible determinant)

Question 9

What is the best kind of Ab to use from immunohistochemistry?

Answer 9

one that can “see” its epitope also when the protein is denatured