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. Other responses: -------------- 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. Best regards, Olindo Assis Chagas'disease Laboratory Centro de Pesquisa René Rachou Oswaldo Cruz Foundation Brasil -------------- 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. Delynn Moss Centers for Disease Control Parasitic Diseases Division
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