Rat IL-6 ELISA or Bioassay

[Michael Dobson]

In article <3lhe12$kfc at yama.mcc.ac.uk>,
Steve Hopkins <shopkins at fs1.ho.man.ac.uk> wrote:
>In article <3lctg2$18a at harbinger.cc.monash.edu.au>, 
>cyberdoc at silas.cc.monash.edu.au says...

>>Could anyone help us out with some info on either:
>>a RAT IL-6 ELISA kit or reagents (including hybridomas)
>>or a RAT IL-6 BIOASSAY (availability of cell lines etc)

>You'll have trouble identifying reagents for an ELISA (though love to 
>hear from anyone who has some): we've been interested in anti-rat 
>cytokine antibodies for ages and they are pretty thin on the ground. 
>Bioassay is easy though. Several line are available. We've used the B9 
>hybridoma (from Luciene Aarden, CLB, Amsterdam) for many years to assay 
>rat IL-6. It is one of the best and most sensitive cell lines for 
>cytokine bioassay and should be easily available. Does recognise murine 
>IL-4 and some neuropoietins, but not well. 
>Let me know if you have a problem getting hold of it.

The monoclonal mouse anti-human IL-6 product from Genzyme shows cross
reactivity with both guinea pig and rat IL-6.  Their polyclonal rabbit
anti-human IL-6 preparation reacts with canine, bovine and chicken IL-6 but
has not been tested on other species, it may work for rat IL-6.  The
polyclonal rabbit anti-mouse IL-6 sera has not been tested against other
species according to the catalog.  You could contact them to see if they
have now done the tests against rat IL-6.

You could also perform the tests yourself.  One way would be to compare an
ELISA with the B9 assay using spleen cells stimulated in vitro.  Incubation
of isolated splenocytes for 24hr in the presence of Concanavlin A produces
levels of IL-6 in the ng/ml range in the culture supernatant.  See
Kalechman et al (1995) Blood 85:1555-1561.  Your control would be
recombinant human IL-6 for the ELISA made from the above reagents.  I
wouldn't try any of the commercial IL-6 ELISA kits for either human or
mouse, the chances of success with rat are not very good.

Mike

-- 
LCDR M. Dobson               | NetNews Admin info.usuhs.mil
Experimental Hematology Dept | dobson at info.usuhs.mil
Armed Forces Radiobiology    |
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