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Immunocytochemistry (ICC) to detect protein in cells (CAT#: STEM-PET-0005-WXH)

Introduction

Immunocytochemistry (ICC) is a common laboratory technique that is used to anatomically visualize the localization of a specific protein or antigen in cells by use of a specific primary antibody that binds to it. The primary antibody allows visualization of the protein under a fluorescence microscope when it is bound by a secondary antibody that has a conjugated fluorophore. ICC allows researchers to evaluate whether or not cells in a particular sample express the antigen in question. In cases where an immunopositive signal is found, ICC also allows researchers to determine which sub-cellular compartments are expressing the antigen.<br />Immunocytochemistry differs from immunohistochemistry in that the former is performed on samples of intact cells that have had most of their surrounding extracellular matrix removed. In contrast, immunohistochemical samples are sections of biological tissue, where each cell is surrounded by tissue architecture and other cells normally found in the intact tissue.




Principle

ICC is dependent on specific epitope-antibody interactions.

Applications

Assess the presence of a specific protein or antigen in cells (cultured cells, cell suspensions).

Procedure

1. Permeabilize Membrane (Optional if detecting a membrane protein)
2. Blocking
3. Primary Antibody
4. Secondary antibody and detection
5. Optional counterstain
6. Cover slips