Storage 

Store at 4℃ and protect from light. When stored as directed, product is stable for at least 6 months.

Description 

The 1× Tyramide Amplification Buffer is designed for cell and tissue immunostaining with tyramide signal amplification. Simply dilute the tyramide compounds in this buffer to make a working solution.

Protocol 

The following protocol is optimized for fixed cells in 96-well plates or tissue sections about 10 mm × 10 mm in size, which can be scaled according to the size of the sample. 

1. According to the sample and antibody, perform the corresponding steps of fixing, blocking and antibody labeling. The sample must be labeled with antibody or streptavidin-HRP conjugates. We recommend to use the UE tyramide signal amplification kit for optimal blocking and secondary antibody labeling. 

2. After labeling the antibody-HRP conjugates, wash the sample 3 times with wash buffer 1×PBS for 5 min each time at room temperature. 

3. Use 1×Tyramide Amplification Buffer to make the dye-tyramide probe into a working solution. The final concentration of the dye-tyramide probe can be changed according to the specific application. Add 100 μL of working solution for each well on 96-well plate or each tissue section. The working solution can be stored at room temperature, protecting from light, for up to 24 hours.. 

4. Incubate at room temperature for 10 min. Protect from light. 

5. Wash the sample 3 times with wash buffer 1×PBS for 5 minutes each time at room temperature. 

6. Optional: If Biotin-tyramide is used, YF-streptavidin can be used for detection, or streptavidin-HRP can be used, followed by DAB to detect. 

7. Microscope imaging. For the tissue samples on the slides, please cover the coverslips and seal them before microscope imaging.