Dear Dr Jian G Zhang,
My understanding is that maintaining saponin in washing buffers helps keeping
membrane pores open so that unbound antibody molecules wash off efficiently from
the cell and thus it reduces background (see Jacob MC et al. (1992), Cytometry
12:550-58 and Carayon P; et al. (1992), J. Immunol. Methods, 147 : 225-30).
These authors use a lower concentration of Saponin for washing (0.02% compared
to 0.1%) than for permeabilization/antibody incubation.
Hope that helps.
Philippe Poncelet
BioCytex, Marseille (F)
"Jian G Zhang" <Jian.G.Zhang@Unilever.com> le 09/06/2000 14:59:32
Pour : Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
cc : (ccc : MailingList/general/Biocytex)
Objet : Intracellular flow cytometry
Dear All,
Just a quick question regarding washing buffer in intracellular flow cytometric
staining: is it
necessary to use saponin containing buffer to wash the cells after
intracellular antibody labelling?
Many thanks for your help.
Jian G Zhang, PhD
Unilever Research
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Arial;}}
\uc1\pard\f0\fs20 Dear All,\par
\par
Just a quick question regarding washing buffer in intracellular flow cytometric
staining:
is it\par
\par
necessary to use saponin containing buffer to wash the cells after intracellular
antibody labelling?\par
\par
Many thanks for your help.\par
\par
Jian G Zhang, PhD\par
Unilever Research\par
\par
}
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