Fluorescamine

Fluorescamine is a spirocyclic compound that is non-fluorescent. Fluorescamine reacts rapidly with primary amine groups in proteins under alkaline conditions to generate products with strong fluorescence (Ex/Em: 390/475 nm). Fluorescamine can be used to detect amine-containing compounds, including amino acids, peptides, and proteins^[1][2]. In Vitro:1. Reagents and instruments
1.1 Fluorescamine solution: Dissolve Fluorescamine in acetone (recommended concentration 1.08 mM), prepare and use immediately (keep away from light).
1.2 Buffer: 0.05 M phosphate buffer (pH 8.0).
1.3 Standard protein solution: bovine serum albumin (BSA) or target protein, concentration gradient 0-1000 μg/mL.
1.3 Instrument: Fluorescence spectrophotometer (excitation wavelength 390 nm, emission wavelength 475 nm).
2. Operation steps and methods (standard curve method)
2.1 Sample preparation: Dilute the protein sample to be tested to the linear range (recommended 1-1000 μg/mL).
Blank control: buffer + Fluorescamine (no protein).
2.2 Reaction: Take 50 μL of sample or standard and add 150 μL buffer (pH 8.0). Then quickly add 50 μL Fluorescamine solution (96-well plate, shake and mix, react for 1-5 minutes).
2.3 Detection: Measure the fluorescence intensity immediately (delay may cause signal attenuation).
3. Data analysis
Use the fluorescence intensity (Y) of the standard protein to perform a linear regression on the concentration (X) to calculate the concentration of the sample to be tested.
4. Precautions
4.1 Please adjust the concentration of Fluorescamine working solution and incubation time according to actual conditions.
4.2 This product is limited to scientific research by professionals and shall not be used for clinical diagnosis or treatment, food or medicine.
4.3 For your safety and health, please wear a lab coat and disposable gloves during operation.
4.4 The reaction time must be consistent (it is recommended to be controlled within 2 minutes).
4.5 Avoid light operation. Fluorescamine is sensitive to light.
4.6 The reaction is best under alkaline conditions.