DAF-FM Chemical Properties

Melting point:

265 ℃

storage temp. 

-20°C

solubility 

DMSO: >1mg/mL

pka

8.19 ± 0.20, most acidic, temperature: 25 °C; 4.23 ± 0.20, most basic(at 25℃)

form 

Solid

color 

clear colorless to yellow

Appearance

yellow to brown solid

λmax

487 nm, 495 nm

Biological Applications

Nitric oxide indicator; detecting microorganisms; measuring Ga i-coupled or Gao-coupled receptors activity

InChI

1S/C21H14F2N2O5/c1-25-13-3-2-8-18(19(13)24)20(28)30-21(8)9-4-11(22)14(26)6-16(9)29-17-7-15(27)12(23)5-10(17)21/h2-7,25-27H,24H2,1H3

InChIKey

DIJCILWNOLHJCG-UHFFFAOYSA-N

SMILES

Fc1cc2c(cc1O)Oc3c(cc(c(c3)O)F)C42OC(=O)c5c4ccc(c5N)NC

Safety Information

Hazard Codes 

Xn

Risk Statements 

22

WGK Germany 

3

Storage Class

11 - Combustible Solids

DAF-FM Usage And Synthesis

Description

DAF-FM (4-amino-5-methylamino-2′,7′-difluorofluorescein) is a cell-impermeant, fluorescent probe for detecting and quantifying low concentrations of nitric oxide (NO). DAF-FM does not need to be activated by cytosolic enzymes and is suitable to detect NO in extracellular matrix.

The fluorescence quantum yield of DAF-FM is ~0.005, but it increases about 160-fold to ~0.81 after reacting with NO and forming a fluorescent benzotriazole (excitation/emission maxima at 495/515 nm) .

The NO detection limit of DAF-FM (~3 nM) is more sensitive than that of DAF-2 (~5 nM). The fluorescence of the NO adduct of DAF-FM is independent of pH above pH 5.5. Moreover, the NO adduct of DAF-FM demonstrates a significantly enhanced photostability compared to that of DAF-2, ensuring reliable results and additional time for imaging.

DAF-FM should be dissolved in DMSO and then used to prepare a working solution. Buffers containing bovine serum albumin (BSA) or phenol red can affect the fluorescence and should be used cautiously.

The cell-permeant version of DAF FM — DAF-FM DA is also available.

Uses

DAF-FM solution has been used as a fluorescent probe for detecting nitric oxide (NO) from plant samples.

Uses

DAF-FM is important reagent for quantitating low concentrations of nitric oxide in solution. This compound is essentially nonfluorescent until it reacts with NO to form a fluorescent benzotrizole.

Uses

DAF-FM has been used in the visualization of nitric oxide (NO) in root sections of wheat seedlings. It has also been used to label neural crest (NC) cells.

Biochem/physiol Actions

DAF-FM is a highly sensitive, photo-stable fluorescent probe for the detection of nitric oxide (NO). DAF-2 is the original compound and is widely used, but DAF-FM is more sensitive to nitric oxide (NO) compared with DAF-2. DAF-FM is more photo-stable and less pH sensitive than DAF-2. DAF-FM is not quite as cell-permeable as the diacetate derivative, DAF-FM, but it does not need to be activated by cytosolic enzymes.