5-(Iodoacetamido)fluorescein

Names

[ Name ]:
5-(Iodoacetamido)fluorescein

[Synonym ]:
N-(3',6'-dihydroxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-5-yl)-2-iodoacetamide
4(5)-(Iodoacetamido)fluorescein

Biological Activity

[Description]:

5-IAF is an idoacetamide derivate of fluoresceine.

[Related Catalog]:

Signaling Pathways >> Others >> Others

Research Areas >> Cancer

Dye Reagents

[In Vitro]

8 µM GSH sample solution is derivatized with 8, 80, 200, 400, 800, 1600, 2000, 2500 µM 5-IAF, respectively and analyzed with CE-LIF. The highest signal intensity is obtained with 400 and 800 µM 5-IAF[2].

[Kinase Assay]

HepG2 cells are lysed in 1 mL 2% acetonitrile in water, and transfered to a 1.5 mL centrifuge tube. An additional 200 µL 2% acetonitrile is used to rinse the dish and combined with the cell extract. To measure the free GSH concentration, three aliquots of 80 µL cell extract are immediately transferred to 0.5 mL centrifuge tubes after a quick mix. Then 20 µL 50 µM NAC is added, followed by the addition of 100 µL 400 µM 5-IAF to the mixture. The derivatization is conducted at room temperature in dark for 1 h, after which the mixtures are centrifuged at 9000× g for 5 min to sediment the insoluble proteins. 10 µL supernatant is diluted 100× with water and analyzed immediately by CE-LIF[2].

[References]

[1]. Salvatore Sotgia, et al. Plasma L-Ergothioneine Measurement by High-Performance Liquid Chromatography and Capillary Electrophoresis after a Pre-Column Derivatization with 5-Iodoacetamidofluorescein (5-IAF) and Fluorescence Detection. PLoS One. 2013; 8(7): e70374.

[2]. Yan Wang, et al. Determination of free and protein-bound glutathione in HepG2 cells using capillary electrophoresis with laser-induced fluorescence detection. J Chromatogr A. 2009 Apr 17;1216(16):3533-7.

[Related Small Molecules]

Chemical & Physical Properties

[ Density]:
1.95 g/cm3

[ Boiling Point ]:
788.4ºC at 760 mmHg

[ Melting Point ]:
265-267ºC (dec.)(lit.)

[ Molecular Formula ]:
C₂₂H₁₄INO₆

[ Molecular Weight ]:
515.25400

[ Flash Point ]:
430.6ºC

[ Exact Mass ]:
514.98700

[ PSA ]:
116.84000

[ LogP ]:
4.41510

[ Index of Refraction ]:
1.781

MSDS

Click to get detail MSDS

Safety Information

[ Personal Protective Equipment ]:
Eyeshields;Gloves;type N95 (US);type P1 (EN143) respirator filter

[ Safety Phrases ]:
22-24/25

[ RIDADR ]:
NONH for all modes of transport

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