Subject: Immunofluorescence

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From:"Harclerode, Donna" <>
To:"''" <>
Date:Wed, 22 Sep 1999 09:42:08 -0700

Can anyone tell me where I can buy rat adsorbed isotype specific
fluorescently conjugated secondary antibodies for immunofluorescence?
Many thanks.


Selina Blades
Laboratory Manager/Departmental Safety Officer
Bloomsbury Rheumatology Unit
4th Floor
Arthur Stanley House
40-50 Tottenham Street
United Kingdom

Hi Selina
I am a bit confused. An anti mouse secondary adsorbed to rat would be a
polyclonal antibody.  With isotype specific anti mouse IgG1, or any other
isotype, you would not adsorb it. PharMingen has very nice isotype specific
antibodies in the biotin format that are QC'd by IHC.  I do not know if
their FITC secondaries are tested in IHC applications.  The PE is not
suitable for IHC.  Not all of their secondaries work in IHC so I would call
and check for whatever isotype you need.  They have isotype specific for all
rat and mouse isotypes and some hamster as well.  I have found only a few
antibodies that will not recognize the polyclonal and require the isotype

Jackson Immunoresearch, have anti mouse (or rat) polyclonal adsorbed IgG and
IgM secondaries.  I have found that often that primaries that are IgM  (both
rat and mouse) will not cross react enough with a plain anti IgG secondary.
They have AMCA, FITC, Cy3 and RITC, all  of which work very well for IHC.
The Cy3 is the brightest if you are doing 1 color work.  With multiple
labeling, Cy3 will bleed into both RITC and FITC so it is not  a good choice
for 2 color. I use the whole IgG but they have F(ab')2 also.  I think the
secondaries made in donkey slightly cleaner than the ones in goat, but both
work very well.  Jackson's adsorbed biotin secondaries are my first choice
for most IHC applications. Their tech service is very helpful and their
shipping is very fast.

PO Box 9
West Grove, PA 19390

European distributor for Jackson ImmunoResearch
Stratech Scientific LTD
61-63 Dudley Street
Luton, Beds LU2 ONP
01582 481895

in the US 1 800-848-6227
in Europe Becton Dickinson GmbH
(49) 40 532 84 48 0

Good luck,

Donna Harclerode, HT, (ASCP) HTL, QIHC
Ligand Pharmaceuticals
San Diego, CA

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