biomolecular interactions

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i69
i69's picture
biomolecular interactions

There are many mitochondrial membrane potential fluroscent dyes which comes handy with different unique proerties for the need.Rhodamine family is new in the feild with varied properties which can help even to study protein folding and protein preotein interactions.
On the otherside to measure reactive oxygen species generated through electron transport chain we have H2DFDA or amplex red.
I want to know the problems expected in loading both dyes at a time and recording the spectra.
Is there any interaction between these two dyes?

macbride
macbride's picture
Molecular Probes is an

Molecular Probes is an excellent resource for information on fluorescent dyes and probes. They have spectra available for scads of molecules. You can check out there website here: http://probes.invitrogen.com/ (they were recently acquired by Invitrogen). Check out the Handbook for extensive data on various fluorescent probe families.

Guy Sovak
Guy Sovak's picture
Thats true Molecular Probes

Thats true Molecular Probes has got great products.
I have a question to you regarding the rhodamine and protein folding.
Could you please elaborate on it.
Have you got some examples and/or are you involved in protein folding research?
Guy

Mohan
Mohan's picture
Yups. Sorry for the delay in

Yups. Sorry for the delay in communication regarding question on biomolecular interaction of fluorescent dyes.
I am working on isolated mitochondria wherein I am measuring free radicals generation from each complex and membrane potential.
To avoid time gap and conditions set forth for the expt. I want to know whether any interaction observed among two dyes molecules if we load them togeter in a single expt. Though many infer not to for this protocol.I want to know what can be the outcome of two dyes loaded together.
Ofcourse spectra shifts if two dyes excitation and emission overlaps.so the results fail,in which case we have to do seperately.
I donot understad the interaction and specificity of fluorophores for a particular expt. Say H2DCFDA is used to measure ROS generation- can we expect the oxidation of the dye only through the radicals generated by target in point. or even from endogeneous resources.
An other question on quantification of fluorescent signals, Can we calculate the activity of target proteins in disceet amounts. If not, why?
What are the draw backs?
Please do not give me references 'cuz i have read books on these topics. I would be glad if anyone can write briefly about it.