A fluorescamine assay was performed to quantify the incorporation of ethylenediamine on the glucan particles. Ethylenediamine-modified particles were dissolved in DMSO to make final concentrations of 0 – 1 mg/mL. A standard curve was prepared using known concentrations of ethylenediamine. The samples or standards (75 μL) were incubated with 25 μL of a 3 mg/mL fluorescamine solution (Sigma, St Louis, MO) for 15 min at room temperature. Fluorescence was measured at EX/EM = 380 nm/ 470 nm using a microplate reader (Tecan). The percentage of incorporation was calculated based on the standard curve.
Fluorescamine solution
Manufactured by Merck Group
Fluorescamine solution is a laboratory reagent used for the detection and quantification of primary amines, including amino acids, peptides, and proteins. It reacts with primary amines to produce a fluorescent derivative that can be detected and measured using spectrofluorometric techniques.
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Lab products found in correlation
2 protocols using fluorescamine solution
1
Quantification of Ethylenediamine-Modified Glucan Particles
2
Quantifying Peptides on Liposomes
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The amount of peptides on liposomes was assayed using fluorescamine, a heterocyclic dione that reacts with primary amines of peptides38 . Specifically, an aliquot (32 μL) of fluorescamine solution (Sigma, cat. No. F-9015; 3 mg/mL in acetonitrile) was added to 100 μL of peptide-conjugated liposomes, and the mixture was allowed to react at room temperature for 10 min. The fluorescence intensity of fluorescamine-labeled liposomes was assessed at an excitation wavelength of 365 nm and an emission wavelength of 470 nm using a SpectraMAX M5 Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA, USA). For calculation of peptides on liposomes, a standard curve was generated using serial dilutions of a cys-promelittin standard solution. The amount of phospholipids in liposomes was determined using a phosphate assay as previously described39 (link).
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