One thing you (and others) might be able to routinely do is to first do staining with fluorescein annexin V, followed by R-PE anti-fluorescein for the apoptotic population. R-PE anti-fluorescein both quenches the fluorescein fluorescence in the green-fluorescence channel (BD FL1) and gives the full R-PE signal in the red-orange channel (BD FL2). Then, after washing off any excess R-PE anti-fluorescein use your green-fluorescent fluorescein-labeled anti-neutrophil antibody. Beyond switching color, the R-PE anti-fluorescein also usually gives an amplified signal that is higher than when using an R-PE direct conjugate. An advantage is that you do not have to buy a new anti-neutrophil antibody. http://www.probes.com/servlets/product?region=Select&item=21250 We have not done this specifically with fluorescein annexin V but I expect that this should work there too. We have done similar "color shifting" and signal amplification for multicolor imaging using Alexa Fluor 594 anti-fluorescein (spectra similar to those of Texas Red). It may be useful for flow if the accessibility of R-PE anti-fluorescein to fluorescein annexin V is poor, but it is not well excited at 488 nm. http://www.probes.com/servlets/product?region=Select&item=11091 Ruth_Wilkins@hc-sc.gc.ca wrote: > Does anyone know of a human antibody for neutrophils that is available in any > colour besides FITC? I am trying to measure Annexin labelling in neutrophils > and am having problems with Annexin-PE but Annexin-FITC works well. > > Thanks, > > Ruth Wilkins, Ph.D. > Consumer and Clinical Radiation Protection Bureau > Health Canada > 775 Brookfield Road, PL 6303B > Ottawa, ON K1A 1C1 > > (613) 941-7263 > FAX (613) 941-1734
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