RE: Intracellular ifn-gamma and IL-4/5

From: Hodge, Greg (HAEM) (hodgeg@mail.wch.sa.gov.au)
Date: Sun Oct 22 2000 - 16:57:21 EST


> ----------
> From:		andrew.head@covance.com
> Sent:		Saturday, 21 October 2000 4:43
> To:	Cytometry Mailing List
> Subject:	Intracellular ifn-gamma and IL-4/5
>
> On the 21st October, Andy wrote:
>      Dear flowers
>
>      I am trying to type Th1 and Th2 cells in Cynolmolgus macaque
>      lymphocytes using intracellular IFN-gamma, IL-4 and IL-5
>      Does anyone have any experience of this ?
>      Using PMA activation(5 hours plus monensin) I have had some success
>      with IFN-gamma, but no luck with IL-4 or 5.Am I supposed to use a
>      different activator for these two monoclonals to work? or incubate
> for
>      a different length of time?
>      Any suggestions would be appreciated.
>
>      Thanks
>      Andy
> --------------------------------------------------------------------------
> ---------------
>
>   Andy,
>	I havent worked with Cynolmolgus macaque lymphocytes, but in the
> case of human lymphocytes you will get a better signal following 24 h
> incubation with PMA (25 ng/ml) + Ionomycin (1ug/ml) (with 1 ug/ml BA or
> Mon Golgi block), especially for IL-4, IL-5.
> Hope this helps.
>
> Regards,
>
>		Greg Hodge PhD
>		email: hodgeg@mail.wch.sa.gov.au
>
>
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