Its principle relies on TSA (Tyramide Signal Amplification) technology, an enzyme-catalyzed immunoassay signal amplification method. The core lies in using HRP (Horseradish Peroxidase) to catalyze the deposition of tyramide derivatives, enabling efficient enhancement of target molecule signals. Through a cyclic staining strategy, it breaks through the target number limitation of traditional IHC/IF, achieving accurate, efficient, and multiplex labeling of multiple target proteins in cell/tissue samples.

Core Principle

1 Specific Target Binding: HRP-conjugated specific antibodies bind specifically to target molecules in samples, forming a "target molecule-primary antibody-HRP-labeled secondary antibody" complex.

2 Enzymatic Activation & Fluorescence Deposition: Add tyramide derivatives labeled with fluorescent/chromogenic groups (Tyramide Conjugate) to the system. In the presence of hydrogen peroxide (H2O2), HRP catalyzes the oxidation of tyramide derivatives to generate highly active tyramide free radicals.

3 Signal Amplification: Active tyramide free radicals rapidly bind covalently to surrounding proteins (mainly tyrosine residues near target molecules), achieving "in-situ deposition" of a large number of fluorescent/chromogenic molecules at target sites. This enables efficient signal amplification and significantly improves the detection sensitivity of target proteins.

Cyclic Staining for Multiplex Fluorescence Labeling

1 Antibody Stripping & Signal Retention: After completing staining and signal amplification of the first target protein, the primary antibody (including antibody complexes) that binds non-covalently to the target and HRP polymer is specifically stripped via antibody stripping. However, the tyramide-fluorophore complex previously covalently linked to tissue proteins remains stable, with sustained fluorescence signals.

2 Multiplex Target Cyclic Detection: For the next target protein to be detected, repeat the above process of "specific antibody binding → TSA signal amplification → antibody stripping". Each cycle uses tyramide derivatives labeled with different fluorophores to ensure distinguishable fluorescence signals for each target.

3 Optimized Detection Sequence: Based on the epitope characteristics (e.g., stability, abundance) of different target proteins, reasonably plan the detection sequence to avoid interference of subsequent staining with already labeled epitopes, ensuring detection specificity and signal quality for each target.

Core Product Advantages

1. Outstanding Detection Performance: High Specificity, High Sensitivity, Low Background

Adopting polymer enzyme-labeling technology, secondary antibodies are bound using polymers as carrier chains, which offers two core advantages:

Enhanced Detection Efficiency: The carrier chain can label multiple HRP molecules, significantly improving detection specificity and sensitivity. The signal amplification factor reaches 10-1000 times, enabling detection of low-abundance target molecules (hard to detect via traditional methods). Meanwhile, signals are only deposited at target sites where HRP acts, reducing background interference and ensuring more accurate results.

Species Limitation Breakthrough: Can bind rabbit- and mouse-derived primary antibodies simultaneously, realizing "rabbit/mouse universality" with stronger adaptability.

Strong Compatibility: Compatible with multiple detection technologies such as immunofluorescence (IF) and immunohistochemistry (IHC).

2. Optimized Experimental Process: Ultra-Simple Operation, Doubled Efficiency

Through technology integration, traditional experimental steps are greatly simplified, reducing operation thresholds and time costs:

Enzyme-Labeled Secondary Antibody Simplification: Compared with ABC method and SP three-step method, no endogenous biotin labeling is required, with more streamlined steps.

Integrated Dewaxing & Retrieval: Eliminates fume hoods and traditional dewaxing/retrieval tanks; integrates three steps ("three-time xylene dewaxing, three to five-time gradient ethanol hydration, antigen retrieval") into a single-solution operation for more efficient processes.

Integrated Staining & Mounting: Combines DAPI staining and anti-fade mounting functions into one solution, reducing reagent replacement and operation steps.

3. Upgraded Detection Capability: Supports Multiplex & Multicolor Labeling, Flexible Adaptation

Meets the demand for simultaneous detection of multiple indicators and adapts to different experimental designs:

One-Stop Multicolor Detection: Equipped with multiple fluorophore-labeled Tyramide (for target protein staining) and DAPI (for nuclear staining), enabling simultaneous multicolor detection of multiple indicators and intuitive presentation of complex sample information.

No Species Restriction for Primary Antibodies: Matched with HRP multi-polymer anti-rabbit/mouse secondary antibodies, primary antibodies can be freely selected from rabbit or mouse sources without being limited to a single species, allowing more flexible experimental design.

Fluorophore Combination for the Kit：

References：

Title：Pan-cancer spatially resolved single-cell analysis reveals the crosstalk between cancer-associated fibroblasts and tumor microenvironment(Quadruple-labeling with Five Colors)

Journal：Molecular Cancer ( IF:37.3)

Cardiac-specific deletion of BRG1 ameliorates ventricular arrhythmia in mice with myocardial infarction（Triple-labeling with Four Colors）

Journal：Acta Pharmacologica Sinica （IF 8.2）

SUMOylated IL-33 in the nucleus stabilizes the transcription factor IRF1 in hepatocellular carcinoma cells to promote immune escape（Triple-labeling with Four Colors）

Journal：Science Signaling（IF 9.5）

Pulmonary interleukin 1 beta/serum amyloid A3 axis promotes lung metastasis of hepatocellular carcinoma by facilitating the pre-metastatic niche formation （Dual-labeling with Three Colors）

Journal：JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH（IF 11.3）