SuperBoost™ Tyramide Signal Amplification is the ideal solution for detecting multiplex fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC) and in situ The most sensitive method for low-abundance targets in hybridization (ISH) applications. SuperBoost kits combine the brightness of AlexaFluor™ dyes with the excellent signal amplification of poly-HRP-mediated tyramine labeling reactions, with sensitivity 10-200 times equivalent to standard methods. The SuperBoost kit is also 2-10 times more sensitive than conventional tyramine amplification technologies such as TSA™. For superior research results, SuperBoost kits provide sharper definition of critical areas that imaging methods struggle to provide.

SuperBoost kits are easy to use and can be easily and flexibly adjusted according to standard ICC, IHC or FISH experimental protocols and are suitable for any type of cells or tissues. Cells labeled with the SuperBoost kit are compatible with any type of microscope, producing high-resolution images for multiplex analysis. This particular kit uses AlexaFluor 647 Tyramine (650/688 excitation/emission wavelengths), which detects through standard Deep Red/Cy5 filter cubes. The kit also contains polyHRP-conjugated goat anti-mouse IgG secondary antibody.

SuperBoost kit features include:
• Ultra-high sensitivity detection of low-level or complex targets through fluorescence imaging technology
• Simple protocol and detection process using standard filters
• Suitable for high-resolution multiplex analysis images co-labeled with DAPI, secondary antibodies or other SuperBoost kits
• Compared to standard ICC/IHC/ISH experiments, the amount of primary antibody required is reduced by 10-100 times< br>
SuperBoost kit is based on the tyramine signal amplification system and utilizes the catalytic activity of horseradish peroxidase (HRP) to label target proteins or nucleic acid sequences in situ at high density. Ordinary ICC/IHC/ISH experiments using SuperBoost kits require 10-100 times less primary antibody than standard ICC/IHC/ISH experiments. SuperBoost kits provide specific signal intensities well above background, allowing protocols to be easily optimized to detect specific signals from samples where high endogenous autofluorescence is observed

Advantages of the SuperBoost Kit

Using Alexa Fluor Tyramine to Enhance Signal: The SuperBoost Kit utilizes Alexa Fluor amides that react with HRP, resulting in bright, photostable Alexa Fluor dye deposits on surrounding proteins and other similar molecules. The SuperBoost kit is the only kit that combines the brightness of Alexa Fluor dyes with the enhanced power of tyramine signal amplification to generate superior signals.

PolyHRP Advantages: Unlike TSA, the SuperBoost kit uses polyHRP-conjugated secondary antibodies. In this system, multiple HRP enzymes are coupled to short-chain polymers, which enhances the signal several times compared to conventional HRP systems. The structural form of poly-HRP enables the antibody to penetrate cells or tissues as effectively as conventional HRP-conjugated secondary antibodies. The average enzyme/antibody protein molar ratio was "4".

Stop Solution: As with any enzyme-based labeling system, signal overflow can occur with this system. The SuperBoost kit contains HRP stop solution to stop the HRP reaction. HRP Stop Solution can be used to obtain the highest possible signal without increasing background signal. The optimized HRP reaction produces images that are as clear as those produced by standard ICC/IHC/ISH methods, but with 10-200 times greater sensitivity.

Lower background: SuperBoost kits contain blockers to eliminate or reduce endogenous peroxidase and fluorescence background signals. These blockers help ensure that only specific signals are enhanced while non-specific/background signals are blocked.