Tyramide Amplification Buffer 

Tyramide Amplification Buffer is a ready-to-use buffer primarily intended for immunostaining of cells and tissues by the technique of tyramide signal amplification (TSA). Tyramide Amplification Buffer enables the reaction between tyramide-labeled antibodies and fluorescently labeled tyramide substitutes (tyramide) using peroxidase, resulting in a highly amplified fluorescent signal. Tyramide Amplification Buffer can be used for signal enhancement in detection methods such as immunofluorescence (IF), immunohistochemistry (IHC), or in situ hybridization (FISH)^[1].

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
The following scheme is applicable to fixed cells in a 96-well plate or tissue sections of approximately 10 mm × 10 mm. It can be scaled proportionally according to the size of the sample ^[1].
Perform the corresponding fixation, blocking and antibody labeling steps using the samples and antibodies. The samples must be labeled with antibodies or HRP conjugates coupled with streptavidin.
After the antibody (streptavidin)-HRP conjugate is labeled, the sample is washed three times with the 1×PBS washing buffer, for 5 minutes each time.
3. Prepare the working solution of the dye-tyramide probe using 1× Tyramide Amplification Buffer. The final concentration of the dye-tyramide probe can be adjusted according to specific applications. Add 100 μL of the working solution to each well of a 96-well plate or to each tissue section. The working solution can be stored under light protection at room temperature for up to 24 hours.
4. Incubate at room temperature in the dark for 10 minutes.
5. Wash the samples three times with the washing buffer 1×PBS, for 5 minutes each time.
6. Optional: If using Biotin-acylamide, fluorescence staining can be performed using fluorescently-labeled streptavidin, or streptavidin labeled with HRP can be used and then stained with DAB.
7. Microscopy imaging. For tissue samples on the slide, cover the slide with a cover glass and seal it, then perform microscopy imaging.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Liquid

Colorless to light yellow

[Tyramide Amplification Buffer]

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

• [1]. Faget, Lauren, et al. "Tyramide signal amplification for immunofluorescent enhancement." ELISA: Methods and protocols. New York, NY: Springer New York, 2015. 161-172.  [Content Brief]