IF750-Tyramide

Tyramide signal amplification (TSA) is a type of enzymatic detection method that uses HRP to perform high-density in-situ labeling of target antigens and can be applied to signal amplification of IF/IHC. The main principles of TSA technology: In the presence of hydrogen peroxide H2O2, horseradish peroxidase HRP can catalyze tyramine as a very active short-lived intermediate. In a short period of time, the intermediate can covalently bind to the electron-rich surface of surrounding proteins to form tyramine complex, and a large number of rich clusters like "island" are integrated with the enzyme as the center. A large amount of luciferin was deposited at the antigen-antibody binding site to achieve signal amplification.

TSA technique can be used to detect low abundance targets that cannot be detected by conventional methods. Signal amplification technology based on tyramine can provide extremely high sensitivity and detect very small amounts of target antigens. TSA technology greatly reduces the amount of antibodies and saves antibodies, and can be used in combination with conventional staining methods for multi-color imaging, or two or more tyramine reactions can be sequentially performed to label different targets on a sample, so multiple fluorescence staining can be achieved with repeated labeling of different fluorescent tyramines

Storage and Handling Conditions

Wet ice packs for transportation, store at -20℃ for 12 months.

Note:

1. After melting at room temperature, the tube is centrifuged for a short time before use.

2. Compared to fluorescent secondary antibody, TSA kit has higher sensitivity and stronger signal. Therefore, the concentration of primary antibody should be reduced, generally expanding 5-10 times on the basis of the dilution ratio recommended in the antibody manual to reduce the background fluorescence caused by non-specific binding. It is recommended to set a resistance gradient concentration for best results.

3. If the background fluorescence is strong, it is recommended to increase the quenching step of tissue autofluorescence.

4. The recommended dilution ratio of fluorescent Tyramide is 1:500, and the dilution ratio can be adjusted according to the experimental results (the recommended dilution ratio range is 1:200-1:1000).

5. Diluent should be prepared by yourself, 1×TBST (G0004 recommended) containing 0.003% H2O2. 1×TBST (G0004 recommended) 100 mL, add 100 μL 3% H2O2, mix well. The diluent is easy to fail, so it is recommended to use it now.

6. If multiple fluorescent labeling is performed, it is recommended to incubate polyclonal antibodies first and then monoclonal antibodies. The antibody corresponding to the high-abundance target protein is incubated first, and then the antibody corresponding to the low-abundance target protein is incubated. For specific operation steps, please refer to our TSAPLus fluorescent multi-label (1-7 labels, 2-8 color fluorescence) dyeing kit (G1226, G1236, G1255, G1256, G1257, G1259).

7. Wear a lab coat and disposable gloves during operation.

8. According to the light source map of the fluorescence imaging system, different fluorescent dye combinations can be selected to carry out TSA fluorescence multi-labeling.

For Research Use Only!