You might try looking at a recently published paper that addresses some of
these issues:
Miyawaki A, et al. Dynamic and quantitative Ca2+ measurements using
improved cameleons. Proc. Natl. Acad. Sci. USA. 96: 2135-2140 (1999).
Trent Watkins
Research Associate / Koshland Lab
University of California at Berkeley
On 14 Apr 1999, EMMA STAVROPOULOS wrote:
>> I'm expressing a conjugate containing ECFP and EYFP from
> 'yellowcam-2' ( Miyawaki et al.,(1997)), and I have found that
> the EYFP fluorescence intensity is considerably lower than reported
> in the paper. This may be due to my particular construct, which
> contains a different binding region to the original. However, I have
> heard reports that EYFP intensity in original constructs varies
> with pH and ion concentration. Can anybody support this? If so,
> what conditions gain optimal EYFP intensity?
>> I would greatly appreciate any advice or comment from anyone using
> these 'Cameleons', in ANY system.
>> Thank-you,
>> Emma Stavropoulos.
>>> e:mail: Emma.Stavropoulos at ed.ac.uk>> Emma Stavropoulos
> Institute of Cell and Molecular Biology
> University of Edinburgh
>> Tel (+44) (0)131 6505319
> Fax (+44) (0)131 650 5392
>>>>>>>> Emma Stavropoulos
> Institute of Cell and Molecular Biology
> University of Edinburgh
>> Tel 0131 6505319
>>
