CPRUSSIN at niaid.nih.gov
Wed Feb 27 10:06:39 EST 2002
Here is a listing of the responses I got.
Tom Nutman's lab here at NIAID does a lot of IgG and IgE ELISAs and they
swear by a company called "Hybridoma Reagent Laboratory". I called them up
and was impressed by the knowledge of their PhD immmunochemist, Roger
Gordon. The company is affiliated with Bob Hamilton, who runs the well
regarded DASY lab at Johns-Hopkins. They specialize in mAbs to human Ig
isotypes and have a huge assortment to choose from. Another plus: very good
prices $110/0.5 mg of biotinylated mAb.
A minus: No fluorochrome labeled mAbs.
At any rate I am going to try these guys.
We use a CALTAG mouse anti-human IgG4 Fc monoclonal conjugated to biotin.
Product number MH1542. We use it to detect binding of an anti-human CD33
monoclonal that started life as a mouse IgG1 but was engineered into a human
IgG4 for use in clinical trials. This Caltag product is fine for us, but we
haven't compared it to anything else.
We have used a biotinilated anti-human IgG4 from SIGMA (clone
8c/6-39, cat # B 6775, lot 118H4885) diluted 1:800 in PBS 10% FBS with
simultaneous incubation with SAPE (Southern Biotechnology Associates cat#
7100-09) diluted 1:800 in PBS 10% FBS with good results.
We have tested this antibody on a checkboard protocol to identify
any cross-reactivity with IgG1, G2, G3, IgA, IgE and IgM. All results were
negatives, indicating the good quality of the reagent. We have had some
problems with cross-reactivity in the past.
I hope it helps.
Centro de Pesquisa René Rachou
Oswaldo Cruz Foundation
Zymed in California (800-874-4494) has biotinylated (and possibly
fluorochrome conjugated) mouse monoclonal antibodies to human IgG4 that work
great on immunoblots. I have not used them in flow, but if I had a need,
this would be my first choice.
Centers for Disease Control
Parasitic Diseases Division
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