Immunocytochemistry: Difference between revisions
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= | =Introduction= | ||
Immunocytochemistry (ICC) is the process of taking tissue culture cells and applying antibodies to determine the spatial and temporal features of a particular antigen (often protein). A survey of the literature reveals many different techniques. However, there is often very little comment about the individual steps of the techniques, and hopefully this page will lead to a productive discussion. Often different techniques applied to the same antibody will reveal different results. | Immunocytochemistry (ICC) is the process of taking tissue culture cells and applying antibodies to determine the spatial and temporal features of a particular antigen (often protein). A survey of the literature reveals many different techniques. However, there is often very little comment about the individual steps of the techniques, and hopefully this page will lead to a productive discussion. Often different techniques applied to the same antibody will reveal different results. | ||
Immunofluorescence (IF) is a subset of ICC in which the antibody localization is visualized by fluorescence. | Immunofluorescence (IF) is a subset of ICC in which the antibody localization is visualized by fluorescence. The below protocol is focused on indirect IF (the use of a secondary antibody). | ||
=The Seven Basic Steps= | |||
# Grow Cells | |||
# Fix Cells | |||
# Permabalize Cells | |||
# Block Cells | |||
# Incubate with Primary Antibody | |||
# Incubate with Secondary Antibody | |||
# Place on a slide | |||
==Grow Cells== | |||
The key point to be made here is to not have too many or too few cells on the day you harvest the cells. |
Revision as of 04:48, 14 June 2006
Introduction
Immunocytochemistry (ICC) is the process of taking tissue culture cells and applying antibodies to determine the spatial and temporal features of a particular antigen (often protein). A survey of the literature reveals many different techniques. However, there is often very little comment about the individual steps of the techniques, and hopefully this page will lead to a productive discussion. Often different techniques applied to the same antibody will reveal different results.
Immunofluorescence (IF) is a subset of ICC in which the antibody localization is visualized by fluorescence. The below protocol is focused on indirect IF (the use of a secondary antibody).
The Seven Basic Steps
- Grow Cells
- Fix Cells
- Permabalize Cells
- Block Cells
- Incubate with Primary Antibody
- Incubate with Secondary Antibody
- Place on a slide
Grow Cells
The key point to be made here is to not have too many or too few cells on the day you harvest the cells.