iF488-Tyramide, 50 μL (for Fluorescent TSA)

Product Information

Description

Tyramide signal amplification (TSA), a class of enzymatic assays that utilize HRP for high-density in situ labeling of target antigens, can be applied to signal amplification of IF/IHC. The main principle of TSA technology: In the presence of hydrogen peroxide (H₂O₂), horseradish peroxidase (HRP) can catalyze tyramine into a very active short-lived intermediate, in a short time, the intermediate can be covalently bound to the electron-rich surfaces of the surrounding proteins to form a tyramine complex, a large number of enriched into the enzyme-centered "island"-like polygroups, and the use of fluorescent dyes labeled tyramine to form large amounts of fluorescein deposits at the antigen-antibody binding site to achieve signal amplification. The enzyme centers on an "island"-like aggregation group, and the use of fluorescent dye-labeled tyramine results in the formation of a large number of fluorescein deposits at the antigen-antibody binding site, thus signal amplification is achieved.

Tyramine-based signal amplification can be used to detect low-abundance targets that cannot be detected by conventional methods. Tyramine-based signal amplification provides excellent sensitivity and detection of very small amounts of the target antigen. Tyramine signal amplification technology greatly reduces the amount of antibody used and saves antibody. The Tyramine Signal Amplification Kit can be used in conjunction with conventional staining methods for multicolor imaging, or two or more tyramine reactions can be performed sequentially to label different targets on a single sample, thus allowing multiple fluorescent stains to be achieved using repeated labeling with different fluorescent tyramines.

Storage and Handling Conditions

Ship with wet ice, store at -20, valid for 12 months.

Component

Note:

1. After the product is melted at room temperature, centrifuge it briefly, blow and mix it with a clean suction head, then take it out.

2. Compared with fluorescent secondary antibody, TSA kit has higher sensitivity and stronger signal. Therefore, the concentration of primary antibody should be reduced, and the dilution recommended by the antibody manual should be increased 5-10 times to reduce the background fluorescence caused by non-specific binding. It is recommended to set the gradient concentration of primary antibody to obtain the best effect.

3. If the background fluorescence is strong, add tissue autofluorescence quenching step.

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 ranges from 1:200 to 1:1000).

5. The diluent to be prepared is 1 x TBST (recommendation G0004) containing 0.003% H₂O₂. 100 mL 1×TBST (recommended G0004), add 100 μL 3% H₂O₂, and mix well. This dilution is prone to failure and is recommended to be used now.

6. If multiple fluorescent labeling is performed, it is recommended to incubate the polyclonal antibody first and then the monoclonal antibody; the antibody corresponding to the high abundance target protein was incubated first, and then the antibody corresponding to the low abundance target protein was incubated. For specific procedures, please refer to our TSAPLus Fluorescent Multi-Label (1-7 markers, 2-8 colors fluorescence) Staining Kit (G1226, G1236, G1255, G1256, G1257).

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

Different fluorescent dye combinations can be selected according to the light source mapping of the fluorescence imaging system to carry out TSA fluorescence multilabeling.

The fluorescence spectral data of the fluorescent dyes associated with the TSA series fluorescent staining kits are as follows:

For Research Use Only!

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