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Cat. No.: TSA555-25 (for 25μl)

Cat. No.: TSA555-100 (for 100μl)

Description

The TSA Tyramide-substrate product line consists of ten tyramide substrates including TSA Tyramide-Biotin (200X), TSA Tyramide-488 (200X), TSA Tyramide-555 (200X) and TSA Tyramide-647 (200X). Relying on tyramide signal amplification technology, these reagents enable immunostaining of cellular and tissue specimens, and are suitable for signal amplification, multiplex fluorescent staining and chromogenic reactions during immunolabeling procedures.

Tyramide Signal Amplification, abbreviated as TSA, is also referred to as Catalyzed Signal Amplification (CSA) or Catalyzed Reporter Deposition (CARD). Thanks to its outstanding signal amplification capacity and compatibility with multiplex staining, this technique is widely adopted in Immunofluorescence (IF), Immunohistochemistry (IHC) and In Situ Hybridization (ISH). It ranks among the most sensitive approaches for detecting ultra-low or low-abundance target antigens in cells and tissues, and markedly improves the precision and sensitivity of target antigen detection without compromising sample resolution.

TSA is a highly sensitive signal amplification technique based on enzymatic catalytic reactions. Its core principle is as follows: horseradish peroxidase (HRP) catalyzes tyramide substrates (e.g., Tyramide-488, Tyramide-Biotin) to generate active tyramide substrates carrying highly reactive free radicals. These free radicals covalently bind to adjacent proteins such as tyrosine residues, depositing massive amounts of labels (specific fluorescent probes, biotin, etc.) within approximately 0.1–1 μm of target sites to achieve cascade signal amplification.

After the first round of primary antibody incubation, secondary antibody incubation and TSA Reagent reaction, antibody complexes non-covalently bound to target antigens can be stripped via enhanced immunostaining antibody elution buffer or heat-induced antigen retrieval, while covalently conjugated fluorescent probes remain stably retained. Subsequent incubation with antibodies from distinct or identical host species can be performed directly without cross-interference from antibodies used in prior rounds to label additional target antigens. This cyclic staining process is repeated until all antibodies and matching tyramide substrates have been applied. All staining signals are then detected collectively, enabling multiplex fluorescent staining on a single specimen.

This product portfolio features a comprehensive selection of ten tyramide substrates, allowing users to select and combine suitable substrates according to the fluorescence channel specifications of their detection instruments. The system delivers exceptional sensitivity, enabling detection of low-abundance target antigens undetectable by conventional staining protocols. It also significantly reduces antibody consumption via tyramide signal amplification. In addition, the reagents exhibit excellent stability, compatible with classic staining workflows for multicolor imaging. The covalently deposited labels resist washout, permitting two or more sequential tyramide reactions to label different biomarkers within one sample.

Storage

Store at -20 °C protected from light; valid for 1 year.

Precautions

• TSA Tyramide-substrate is supplied in a small volume. After thawing, perform a brief spin-down to collect liquid adhering to the tube cap or side walls at the bottom of the tube, then mix thoroughly before use.
• Select appropriate tyramide substrates or multiplex staining kits according to the exact fluorescence channel parameters of your detection equipment. Fluorescence spectral data for all TSA Tyramide-substrates are listed in the appendix. Our tyramide substrates are named based on excitation wavelengths, while comparable products from other suppliers may adopt emission wavelength-based naming conventions; please distinguish between the two naming standards carefully.
• TSA technology delivers superior sensitivity and stronger signal intensity, so the working concentrations of primary and secondary antibodies should be appropriately reduced. Adjust antibody dilution ratios based on experimental outcomes. Generally, antibodies can be further diluted 2–10 fold relative to standard IHC dilution ratios to minimize background fluorescence caused by non-specific binding. Set up serial dilution gradients during initial testing for optimization.
• Photobleaching occurs with all fluorescent dyes. Keep samples shielded from light as much as possible to slow signal fading.
• For frequent short-term use, store the product at 4 °C, where it remains functional for at least 1 to 2 weeks. Repeated freeze-thaw cycles should be avoided as much as possible.
• This product is intended solely for scientific research conducted by qualified professionals. It must not be used for clinical diagnosis or therapy, food or pharmaceutical manufacturing, nor stored in residential spaces.
• For your personal safety and health, wear a lab coat and disposable gloves throughout all handling operations.