Tyramide signal amplification (TSA) technology can be used to detect low-abundance targets in tissue and cellular immunofluorescence (IF), immunochemistry, and in situ hybridization (ISH) experiments. This technology can increase the signal sensitivity by about 100 times. The HRP-labeled secondary antibody can activate the fluorescently labeled tyramide molecules in the presence of H2O2, and convert the labeled tyramide into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues in the vicinity of the reaction site. The reaction is rapid (less than 10 minutes) and the deposited labeling can be observed directly under a standard or confocal microscope. TSA fluorescence kits can also be used in combination with traditional immunofluorescence methods for multicolor imaging, and two or more tyramide reactions can be performed sequentially to label different targets on one sample. The use of TSA reagents significantly improves signal sensitivity while maintaining the specificity and resolution ratio compared to common experiments, in addition, TSA reagents can significantly reduce the consumption of primary antibodies or probes.
The fluorescent molecule of this kit is HyperFluor 488 (495 nm/519 nm), which can be detected by standard FITC filters. Compared to fluorescein in the Fluorescein TSA Fluorescence System Kit (K1050), HyperFluor488 is more photostable, allowing for longer observation and image capture, it also obtains low self-quenching and high fluorescence quantum yield.
Our products Tyramide Signal Amplification (TSA) kits include several kits with different wavelength: K1050 with fluorescein for detection at excitation and emission wavelengths of 494 nm/517 nm, K1051 with Cyanine 3 for detection at excitation and emission wavelengths of 550 nm/570 nm, and K1052 with Cyanine 5 at excitation and emission wavelengths of 648 nm/667 nm. You can purchase TSA kits with different wavelengths according to your requirements.
Schematic diagram of tyramide signal amplification system
Label the cell or tissue sample with the primary and secondary antibodies using conventional methods. The horseradish peroxidase (HRP) conjugated to the second antibody catalyzes the conversion of the labeled tyramide to a reactive radical, and the tyramide radical is covalently bound to a nearby tyrosine residue to provide a high-density label.
