[Cytometry] PE query

Mario Roederer roederer at drmr.com
Mon May 2 07:20:15 EDT 2011


I wouldn't use PE.  Given essentially no details about what you are trying to accomplish, it's virtually impossible to suggest much. But I can't think of an experiment where I would use PE.

If you are studying non-specific endocytosis (pinocytosis), which is about the only way PE would be taken up, you would need to use high concentrations to get signal.  But then the problem is that the proteolytic environment of the early endosome (and lower pH) will start degrading the PE and you will lose signal.  And to use PE for this is phenomenally expensive.  Instead, consider using a conjugated dextran (eg from Sigma, I think), which you can use economically at the nearly mg/ml levels you need for pinocytosis experiments.

As for "all cell types", the answer there is also no.  Every cell type is different.

If you are just trying to stain cells without relying on an active procees, consider using a membrane probe like Nile Red.  Again, very inexpensive and it WILL label all cells.

mr


On May 2, 2011, at 12:48 AM, Jasneet Kaur Phd2010 wrote:

> hi all,
> 
> i have a Phycoerythrin dye in my lab (not conjugated to anything) obtained
> from Molecular Probes. i have to use a dye to just stain a cell population
> with it. does this dye enter/bind to all cell types without any preference?
> also, if possible, can anyone send me the protocol for its use? i am
> completely clueless about the concentration/duration of the treatment.
> thanks in advance.
> 
> -- 
> Jasneet Kaur
> c/o Dr. Satyajit Rath
> Research Scholar
> Immunobiology Lab-2
> National Institute of Immunology,
> New Delhi
> _______________________________________________
> Cytometry mailing list
> Cytometry at lists.purdue.edu
> https://lists.purdue.edu/mailman/listinfo/cytometry
> Search the list archive at  http://tinyurl.com/cytometry




More information about the Cytometry mailing list