Hello Jan, we detect cytokine production in transfected CHOs using paraformadehyde fixation and saponin permeabilisation. If you need the protocol I'll be happy to send it to you. Mara Dr. Mara Rocchi DVM Ph.D. Moredun Research Institute Pentland Science Park Bush Loan, Penicuik EH25 0PZ Scotland, UK Tel: +44-131-4455111 Fax: +44-131-4456111 e-mail: mara.rocchi@mri.sari.ac.uk -----Original Message----- From: Jan Grawé [mailto:jan.grawe@rudbeck.uu.se] Sent: 22 November 2002 09:03 To: cyto-inbox Subject: Cytokine measurement in transfected cell line Dear All, I have a user who wishes to detect cytokine production in a transfected cell line, see below . Use of at least one commercial permeabilisation kit has failed. Literature and email archive searches have yielded virtually no information, so any insights are appreciated! Jan Grawé >Hi, > >I am trying to develop an intracellular staining protocol to detect >intracellular protein in a tumor cell line >by flow cytometry. I have checked the literature and the technique >seems to be used almost entirely for >lymphoid cells. Does this mean that other types of cells are >difficult to permeabilize? > >Does anyone have any experience detecting intracellular protein >within tumor cells using flow cytometry? > >I am transducing the cell line MB49 (murine bladder carcinoma) with >recombinant adenovirus expressing IL-15, >and I would like to detect IL-15 by intracellular staining. > >I would really appreciate any insight and/or comments. > >Thanks, >Christina Ninalga -- -------------------------------- Jan Grawé Cellanalyslab Rudbecklaboratoriet/C5 Uppsala Universitet Dag Hammarskjölds väg 20 75185 Uppsala tel: +(0)18-4714656 mobil:+(0)70-2577874 fax: +(0)18-552739 epost: jan.grawe@rudbeck.uu.se http://www.rudbeck.uu.se/cellanalys The information contained in this e-mail message and any files transmitted with it is confidential and may be legally privileged. It is intended only for the addressee and others authorised to receive it. If you are not the intended recipient or the person responsible for delivering the message to the intended recipient you are advised that you have received the e-mail in error and that any disclosure, copying, distribution or action taken in reliance on the contents of the e-mail and its attachments is strictly prohibited and may be unlawful. Please advise the sender immediately by replying to this e-mail and delete it and all copies from your system.
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