HMBR are not fluorescent by themselves, but when combined with Y-FAST, they emit yellow light when excited by blue light.

Instructions for use
I. Solution preparation
1. Stock solution: The recommended concentration is 10–50 mM (adjustable according to experimental needs). Dissolve it in sterile water or buffer and mix until completely dissolved.
Notes:
Powder: Store at -20°C or lower in a light-proof, dry environment; Solution: Store at 4°C for short term, and store at -20°C for long term in aliquots and light-proof conditions to avoid repeated freezing and thawing.
II. Operation steps (taking oxidation reaction as an example)
1. Prepare the reaction system: buffer (such as 50 mM Tris-HCl, pH 7.4).
2. Add HMBR stock solution to the working concentration (usually 1–5 mM).
3. Add enzymes or other reactants (such as peroxide).
4. Control reaction conditions: Temperature: 37°C. Time: 10–30 minutes, the specific time is optimized according to the experiment.
5. Detection method: Use a spectrophotometer to measure the absorbance or chemical signal of the reaction product.