pharmingen cytokine frustration: DUO solution?

[Ress, S, Stan, Dr]

On 17 october J Mathew Buchanan wrote:

>From: jbuchana at magnus.acs.ohio-state.edu (J Matthew Buchanan)
>Subject: Pharmingen IL-4 Sandwich ELISA - High Background Probs
>..using the Pharmingen sandwich ELISA (11b11 capture, BVD6-24G2
>biotinylated detecting) and am having problems with real high
>background.... I have dealt with Pharmingen's tech support
>and this has only added problems.  They gave me a different lot #
>of both coating and detecting Abs, but the problem remains.


 and similarly from: yarosh at sask.usask.ca

> RE: Pharmingen IL-4 Sandwich ELISA - High Background Probs
>Date: 17 OCT 94 23:08:43 GMT

>We've been working on the sandwich ELISA for both IL-4 and IFN 
>gamma using Pharmingen reagents on Immulon 2 plates. The IFN 
>gamma works no problem but we've
>had to play around with conditions. The IL-4 ELISA gave us high 
>backgrounds
>no matter what we tried and we're getting ready to give up. Although 
>the Pharmingen IL-4 MAbs work great in an ELISPOT... if that's any 
>consolation.

Is this problem specific for murine ELIZA, or does it also occur with 
ab's to human cytokines? As far as I know, Pharmingen don't make 
any "perfect pairs" to detect human IFN-gamma.
Genzyme are now advertising a DUOSET which is more 
economical-does anyone have experience with this product re: 
quality, bg, etc?  Any info would be most appreciated.

Stan.



Stanley Ress
Clinical Immunology laboratory
H47 Old Main Bldg
Groote Schuur Hospital and UCT
Cape Town, 7925
South Africa

TEL:INTERN. + 021-7946176
FAX:   "    + 021-4486815
e-mail:  SRESS at uctgsh1.uct.ac.za