Chromogenic in situ Hybridization (CISH) (CAT#: STEM-MB-1183-WXH)

Introduction

A simple and practical alternative to FISH that does not depend on expensive fluorescence microscopy instrumentation and expertise has been developed and refined since 2000 and is called Chromogenic in situ Hybridization (CISH).<br />In addition to providing a user-friendly economical alternative to FISH, chromogenic hybridization methods have the distinct advantage of enabling visualization of tissue context, including nucleus and cell shape, together with the probe signal on the same image. Unlike the majority of fluorescent detection reagents, chromogenic agents used in most CISH methods are chemically stable and do not fade over time, allowing easy storage and repeated re-examination of samples.

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Principle Applications Procedure Materials Notes Related Products

Principle

In CISH, probes are labeled with an antigenic moiety and detected via antibodies conjugated to an enzyme, typically horseradish peroxidase (HRP) or alkaline phosphatase (AP), that catalyzes reactions of chromogenic substrates. The resulting chromogens precipitate at the probe target site and can be detected under a standard bright-field microscope.

Applications

Assess gene amplification.
Detection of chromosomal rearrangements and fusions.

Procedure

1. Probe design
2. Prerartion of tissue
3. Hybridization of probes
4. Detection

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