5-FAM SE, also called 5-FAM NHS, is consist of two parts, the fluorophore 5-FAM and the reactive group N-Hydroxysuccinimide (NHS).

Store at -20°C in dark

Guidance for conjugation of Fluorophore-NHS with protein: Step 1 Solutions & buffers 1 M Sodium Phosphate Buffer, pH 7.0 10x NaH₂PO₄ 1 M 125 ml Na₂HPO₄ 1 M 875 ml Reaction buffer Sodium Phosphate buffer (pH 7.0) 0.1 M 100 ml KOAc(pH 7.4) 75 mM 37.5 ml DTT 2 mM 2 ml Step 2 Make a 20 mM stock of Fluorophore-NHS (e.g. 5-FAM NHS) in DMSO Step 3 Enzyme storage buffer HEPES–KOH (pH 7.4) 20 mM 20 ml KOAc (pH 7.4) 100 mM 50 ml DTT 2 mM 2 ml Glycerol 10% 100 ml Preparation: Pure lyophilized protein can be dissolved in reaction buffer to a final concentration of >2.0 mg ml^-1. If the protein is already in solution, it should be brought into reaction buffer by dialyzing overnight at 4°C against 2 l reaction buffer, by gel filtration, or by repeated buffer exchange through a protein concentrator of desired MWCO

5-FAM SE, also called 5-FAM NHS, is consist of two parts, the fluorophore 5-FAM and the reactive group N-Hydroxysuccinimide (NHS). 5-FAM is a 5-carboxy derivative of fluorescein. When proteins conjugated with 5-FAM NHS through the reaction of NHS with the amine groups, the proteins are labeled with 5-FAM with excitation wavelength of 492 nm and emission wavelength of 514 nm (PH=9).