M_Doherty at NIH.gov
Mon Jan 22 09:48:45 EST 1996
In article <Pine.3.89.9601190034.A539063573-0100000 at SKYCAT.USask.CA>,
cheol.yun at usask.ca wrote:
> Hi netters!
> Would anyone tell me about proper way of doing cytokine depletion (IFNg and
> IL-4) in mice (C57BL/6, if possible)?
> How much antibodies are required how often?
> Were you actually able to deplete cytokines?
> Any input would be greatly appreciated.
How long do you intend to deplete for? A single shot of 2-3 mg/mouse
(with a good neutralising antibody) leads to a fairly strong transient
depleteion of many cytokines. In our hands it works nicely for both
IFN-gamma (XMG 1.2) and IL-4 (11B11). You get an almost complete
disappearance of ctyokine activity (as measured by T cell subset
development, and antibody isotype switching) and often the protein is
undetectable by ELISA.
For longer treatments, we injected 2 mg/mouse every 3 weeks with
maintainance doses of 1 mg/moue each week. Alzet pumps implanted
subcutaneously worked better for some other anti-cytokine mAb (such as
anti-TNF alpha), which otherwise needed to be given every 24 hours.
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