5-FAMSE is a cell-permeable green fluorescent labeling reagent used for labeling peptides, proteins and nucleotides, demostrating excitation and emission maxima are 494 and 520 nm, respectively. 5-FAM SE can react with amines and can yield stable amine conjugates.

Physicochemical Properties

Molecular Formula	C25H15NO9
Molecular Weight	473.3879
Exact Mass	473.074
CAS #	92557-80-7
Related CAS #	5-FAM;76823-03-5
PubChem CID	2762599
Appearance	Light yellow to yellow solid powder
Density	1.7±0.1 g/cm3
Boiling Point	778.7±70.0 °C at 760 mmHg
Melting Point	>300°C (dec.)
Flash Point	424.7±35.7 °C
Vapour Pressure	0.0±2.8 mmHg at 25°C
Index of Refraction	1.790
LogP	0.71
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	3
Heavy Atom Count	35
Complexity	894
Defined Atom Stereocenter Count	0
InChi Key	GECIDMICWWDIBO-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H15NO9/c27-13-2-5-17-19(10-13)33-20-11-14(28)3-6-18(20)25(17)16-4-1-12(9-15(16)24(32)34-25)23(31)35-26-21(29)7-8-22(26)30/h1-6,9-11,27-28H,7-8H2
Chemical Name	2,5-dioxopyrrolidin-1-yl 3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-5-carboxylate
Synonyms	5-FAMSE 5 FAMSE 5FAMSE CFSE 5-FAM-SE
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

Luminescently released 5 may be quantified with high sensitivity -HT using capillary electrophoresis (CE)-laser-induced fluorescence (LIF) techniques for the measurement of serotonin and 5-FAM SE (5-carboxyfluorescein N-succinimide ester) derivatization. For 5-HT derivatization, CFSE is a great dye [1]. Hydriodic acid (HI)-induced reductive hydrolysis of pheomelanin yields two labeled aminohydroxyphenylalanine (AHP) isomers, which may be detected using 5-FAM SE (5-carboxyfluorescein succinimide ester) as the labeling reagent. Pre-capillary derivatization occurs in the body [2].

References

[1]. Quantification of luminally released serotonin in rat proximal colon by capillary electrophoresis with laser-induced fluorescence detection. Anal Bioanal Chem. 2009 Apr;393(8):2059-66. [2]. Sensitive determination of pheomelanin after 5-carboxyfluorescein succinimidyl ester precapillary derivatization and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jan 1;861(1):136-9.

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~52.81 mM)

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1124 mL	10.5621 mL	21.1242 mL
	5 mM	0.4225 mL	2.1124 mL	4.2248 mL
	10 mM	0.2112 mL	1.0562 mL	2.1124 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.