Does anyone know what is the best buffer to keep on the pH on a medium (complete and KH) with Nigericin and Monensin to permeabilize our fibroblasts and detect through FITC-dextran their intralysosomal pH by flow cytometry? We need keep constant pH to realize our calibration line.
Dear Maria,
It might be not easy to find a short and 100% correct answer to your interesting but complex question.
Perhaps if someone works in this your field he/she will have an reliable opinion and perhaps can / will tell you his or her “optimum buffer system”.
Until there will be a sufficient, valuable and reliable reply by a colleague involved in cell culture experiments of this type I only honestly can guess that it might be a good idea to go through the references I posted below (which I will not comment on here and only provide the URLS which either end up with a pdf – free access- or at least at an abstract page, if the article is available only by PPV = pay per view)
Hopefully you will find – at least rudimentary - an answer or solution for your complex experimental setup,
with my best regards and wishes, Wolfgang
References:
(2013) http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055526
(2012) http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3348458/
(2006) http://digitus.itk.ppke.hu/~cseri/edu/Books/Cell%20Biology/Volume%203/Part%20A/Chapter16.pdf, cf. p.165ff
(2003) http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.10028/pdf
(1998) http://www.jbc.org/content/273/4/2044.full
(1993) http://www.sciencedirect.com/science/article/pii/089158499390039W
(1989) http://www.ncbi.nlm.nih.gov/pmc/articles/PMC303693/pdf/jcinvest00083-0038.pdf
(1986) http://www.sciencedirect.com/science/article/pii/0304416586903259
(1982) http://www.ncbi.nlm.nih.gov/pmc/articles/PMC346883/pdf/pnas00456-0183.pdf
(1982) http://jcb.rupress.org/content/93/1/57.full.pdf
thannnnkkksss soo muchh!!!!!! really!! if finally i can find what is the best for my proces I'll let you know!! thanks one more time!
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