BCECF (2′,7′-Bis(2-carboxyethyl)-5(6)-carboxyfluorescein)

A fluorescent pH probe

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1120.00

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货号: ajce44782
CAS: 85138-49-4
别名: 2,7-双(2-羧乙基)-5(6)-羧基荧光素,2′,7′-Bis(2-carboxyethyl)-5(6)-carboxyfluorescein
分子式: C27H20O11
分子量: 520.44
纯度: >98%
溶解度: DMSO : 62.5 mg/mL (120.09 mM);Water : < 0.1 mg/mL (insoluble)
储存: Store at -20°C, protect from light
库存: 现货

Background

BCECF is a fluorescent probe commonly used to measure pH.¹ It is a dual-excitation ratiometric pH indicator with a pK_a of ~6.98. Measurements of pH are made by determining the ratio of emission intensity, detected at 535 nm, when excited at 490 nm versus the emission intensity when excited at 440 nm.¹ BCECF is membrane impermeable, whereas the acetoxymethyl ester, BCEBF-AM is membrane permeant.

1.O'Connor, N., and Silver, R.B.Ratio imaging: Practical considerations for measuring intracellular Ca2+ and pH in living cellsMethods Cell Biol.81415-433(2007)

Protocol

Kinase experiment:

The cells are palced in a petri dish and BCECF present in trichomonads is fixed for electron microsopy. Controns without BCECF loading, without DAB treatment, and without UV illumination are run in parallel. Trichomonads remaine viable during the procedure, except that without BCECF they are dead and disrupted by 4 h. Then the cell are lysed. After lysis, BCECF is added to the lysate to 20 μM, and the lysate is incubated,followed by 3 washes in lysis buffer to remove free dye and resuspension of the pellet in the original volume[1].

参考文献:

[1]. Scott DA, et al. Analysis of the uptake of the fluorescent marker 2',7'-bis-(2-carboxyethyl)-5(and-6)-carboxyfluorescein (BCECF) by hydrogenosomes in Trichomonas vaginalis. Eur J Cell Biol. 1998 Jun;76(2):139-45.