Looks like one option is BCECF/AM--from a paper I was reading recently (Cell
Physiol Biochem. 2008;22(1-4):15-30. Epub 2008 Jul 25.) However, you would
probably want to use the non-permeable variant, if you want to monitor the
endocytosed vesicles.
Jacob
"Cells were loaded with BCECF/AM (Molecular probes) as a probe of cellular pH,
at a loading concentration of 5 ?M for 15-30 min. For BCECF, excitation lights
was set at 430 nm and 490 nm at a rate of 100 Hz and emission was monitored at
540 nm (6 nm band pass). A calibration curve was performed at the end of each
experiment by application of a 100 mM KCl, 10 mM HEPES solution containing
nigericin (5 ?M) and buffered at pH 6.4 or pH 7.8."
*******************************************
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
Dallos Laboratory
F. Searle 1-240
2240 Campus Drive
Evanston IL 60208
lab: 847.491.2438
cel: 773.608.9185
email: j-kell...@northwestern.edu
*******************************************
----- Original Message -----
From: Daniel Jin
To: CCP4BB@JISCMAIL.AC.UK
Sent: Thursday, May 21, 2009 10:59 AM
Subject: [ccp4bb] pH sensitive fluorescent dye
Hi,
Sorry for the off-CCP4 question. Crystallographers are now doing
everything...
We are looking for a fluorescent dye that will be turned on, or
increase signal, when pH goes from neutral to acidic. More specifically, we
would like to monitor the internalization of a protein into the acidic
endosome. There are dyes that will be quenched at low pH. But we want something
that willl do the opposite. Any information will be highly appreciated.
Best,
Daniel
