I agree with Dick Haugland - Trypan Blue has been used by Bassoe et al, to identify internalized vs attached labeled bacteria in a phagocytosis assay. I believe they also used Crystal Violet at one stage - although I have not tried Crystal Violet myself. I believe the first reference to this is "Phagocyte C3-mediated attachment and internalization: flow cytometric studies using a fluorescence quenching technique" Bjerknes,R.; Bassoe,C.- F. Blut 49,315-323, 1984 Paul Robinson --------------------------------------------------------------------------------------------- On 12 Nov 2002, at 6:23, Richard Haugland wrote: > > Does it have a dye on it? If it is labeled with most dyes (except > phycobiliproteins) then trypan blue is an almost universal quencher of > external antibodies (especially fluorescein and Alexa Fluor 488) but not > internalized fluorescent antibodies. Useful trick for answering > "inside-outside" questions. > > > > > > "Barren, Phil" wrote: > > > Could some one point me in the right direction? > > > > I have an investigator that wants to determine if their antibody is > > internalized post binding to the surface of cells. > > > > Could someone offer some advice on detection, controls etc?? > > > > Thank you > > > > Philip Barren J.Paul Robinson, PhD PH:(765)4940757 Professor of Immunopharmacology Professor of Biomedical Engineering Purdue University FAX:(765)4940517 EMAIL:jpr@flowcyt.cyto.purdue.edu WEB: http://www.cyto.purdue.edu
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