Tyramide Signal Amplification

Biotium developed TyraMax™ Amplification Dyes as high-performance TSA dyes, offering brighter, more photostable signals than Aluora® and Opal® reagents. They also remain stable in amplification buffer for up to 24 hours, facilitating automated staining workflows. TyraMax™ Dyes are offered as standalone dye solutions, as 3-color or 5-color dye sets plus DAPI counterstain, and in a sampler for custom panel optimization. See below for alternatives to specific Aluora® and Opal® reagents. Learn more about our reagents for tyramide signal amplification (TSA).

Aluora is a registered trademark of Thermo Fisher Scientific; Opal is a registered trademark of Akoya Biosciences.

Our Tyramide Amplification Kits have been demonstrated to be robust and versatile for multi-color fluorescence imaging, compatible with dye-labeled antibodies and various cell staining methods (see Figure 1).

To use a Tyramide Amplification Kit in addition to one or more dye-labeled antibodies, follow the kit protocol to fix and block samples; label with primary antibodies; then detect primary antibodies using secondary antibodies. Dye labeled secondary antibodies can be co-incubated with the HRP-conjugated secondary or HRP-streptavidin from the tyramide kit. After washing, perform the CF® Dye tyramide reaction according to the kit protocol. The tyramide reaction does not interfere with the binding of dye-labeled antibodies or other fluorescent staining reagents.

Performing multi-color detection with more than one dye tyramide on the same sample requires sequential tyramide staining reactions, followed by HRP inactivation or antibody stripping between each step. See our tech tip:

Multi-Color Fluorescence Imaging Using Biotium’s Tyramide Amplification Kits

Yes, CF® dye tyramides can be used for in situ hybridization using Tyramide Signal Amplification¹ (TSA). They are also compatible with the RNAscope assays  for RISH².

¹ doi: https://doi.org/10.1038/s41598-018-38171-5.

² doi: http://dx.doi.org/10.1101/651083.