In Situ Hybridization Detection Systems†

In Situ Hybridization (ISH) is a powerful technique for detecting and localizing specific nucleic acid sequences within cells or
tissues. This is achieved by the hybridization of a labeled probe to the specific RNA/DNA sequence within the cell and subsequent
detection of the bound probe.

Features

• Enables the semi-quantification of mRNA expression and helps to determine the temporal and spatial patterns of gene expression in cells, tissue and whole animals
• Used for detecting intracellular pathogens with a very high degree of sensitivity

I. SuperSensitiveTM ISH Detection System

This kit is intended for use on cells and tissues from all species (rodent and non-rodent). It is based on the following principle:

1. Cells/tissues are permeabilized and hybridized with a labeled (Digoxin/Digoxigenin or Fluorescein) probe
2. The probe is detected with a probe Hapten-Specific Antibody
3. A Biotinylated secondary antibody may be used (Universal ISH kits with a Biotinylated anti-probe antibody do not require the use of the secondary antibody)
4. The bound antibody is then allowed to react with Streptavidin conjugated to an enzyme marker. Streptavidin binds very strongly to the biotin residues on the anti-probe antibody resulting in the addition of multiple enzyme molecules
5. Finally, a substrate is added that is acted on by the enzyme producing an intense precipitate at the site of hybridization

Benefits

• High specificity inherent to the hapten-antibody and biotin-streptavidin interactions
• High sensitivity due to serial amplification

BioGenex offers a choice of detection points:

AEC (Brick red precipitate)
BCIP/NBT (Purple precipitate)
DAB (Brown precipitate)
Fast Red (Red-colored and fluorescent precipitate)

† All products in this section are for Laboratory Use unless otherwise specified.

ISH Core Kits (Manual Use)

For the convenience of the user, BioGenex offers Core Kits for ISH. These kits provide the user a high degree of flexibility in using a chromogen and a labeled probe of choice.

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