Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4297

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Indirect Detection Using Fluorochrome-Labeled Reagents

Ed Harlow and David Lane

This protocol was adapted from "Staining Cells," Chapter 5, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

The first 100 words of the full text of this article appear below.


INTRODUCTION

Fluorescence-linked detection is best used for high-resolution studies. In addition, some fluorochromes have emission spectra that do not overlap, so two fluorochromes can be observed on the same sample. This allows the study of two different antigens in the same specimen even when they have identical subcellular distributions. Flurorescein, rhodamine, and Texas Red are the most commonly used fluorochromes. Fluorescein emits a yellow-green light that is easily detected, but it is prone to rapid photobleaching. Rhodamine emits a red color and is not as prone to fading as fluorescein, but the rhodamine conjugates are more hydrophobic and therefore yield higher . . . [Full Text of this Article]


MATERIALS

Reagents

Equipment


METHOD


TROUBLESHOOTING


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