In Situ Hybridization Workflow

Due to their high affinity, high sensitivity, and low background, fluorescence- and enzyme-based detection reagents from Vector Laboratories are ideal for in situ hybridization (ISH) applications.

Step 1 Labeling


Label your DNA or RNA probes and oligonucleotides using one-step direct methods, thiol-based indirect labeling, or specific 5' and 3' end tagging techniques.

Carbohydrate Labeling DNA / RNA Labeling

Step 2 Characterization


Confirm probe customization and label incorporation. For example, use the QuantTag Biotin Quantitation kit to determine the number of biotin molecules incorporated in each strand of nucleic acid.

Affinity Binding Matrices Biotin Quantitation

Step 3 Hybridization


Optimize the probe concentration and buffer conditions to ensure effective annealing to complementary sequences.

Slide Preparation

Step 4 Blocking


Reduce or eliminate unwanted background (non-specific) staining on tissue sections and cell preparations using a blocking solution. Non-specific staining can result from endogenous enzyme or tissue elements, including endogenous enzyme activity, presence of Fc receptors, or interactions of detection reagents with tissue or cell proteins and other macromolecules. Choose a blocking solution based on the results of negative control sections.

Blocking Reagents

Step 6 Counterstain and Mount

Counterstain and Mount

Retain and preserve the specific target signal for short-term storage or longer term archiving.

Enzyme Substrates Counterstains and Special Stains Mounting Media