Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa FluorŪ 647 conjugated products with eFluorŪ 660 conjugated products.
Description
The anti-mouse IgD polyclonal goat antiserum reacts with the δ heavy chain of mouse IgD, and is a useful polyclonal activator of B cells (in vitro or in vivo). IgD is expressed by mature peripheral B cells. Unlike most monoclonal antibodies specific for IgD, this goat polyclonal anti-IgD antiserum is a strong activator of B cells via engagement of the IgD receptor. The use of anti-IgM as a polyclonal B cell activator is problematic by virtue of interference by the high levels of circulating serum IgM. This goat polyclonal anti-IgD antiserum is useful for in vivo induction of IgE(1) and IL-4/Th2(2) responses.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This polyclonal goat anti-mouse IgD antiserum has been reported and tested for in vivo activation of mouse B cells, as measured by induction of systemic IgE1 and IL-42.
Applications Tested
This polyclonal goat anti-mouse IgD antiserum has been tested for in vivo activation of mouse B cells, as measured by induction of IgE1 and IL-4 (Th2)2 responses. A dose of 200 ul per mouse is suggested for induction of IgE responses, administered by subcutaneous or intraperitoneal injection. High levels of IgE were measurable by ELISA within 5-7 days of administration. High levels of IL-4 were measurable by in vivo cytokine capture assay within 5 days of administration. Intraveneous injection is not recommended.
Induction of IgE In Vivo: Treatment of 3 mice with 0.2 ml each of the goat anti-mouse IgD antiserum yielded circulating IgE levels of 2.7 μg/ml, 1.9 μg/ml, and 1.3 μg/ml within 8 days; untreated control mice were found to have less than 76 ng/ml circulating IgE.
Induction of IL-4 In Vivo: Treatment of 5 mice with 0.2 ml each of saline control or the goat anti-mouse IgD antiserum yielded IL-4 levels of 96 +/-1.23 pg/ml (geometric mean and standard error) and 3,583 +/- 1.03 pg/ml, respectively, as measured by in vivo IL-4 capture assay.
References
1. Finkelman F, Snapper C, Mountz J, and Katona I. 1987. Polyclonal activation of the murine immune system by a goat antibody to mouse IgD. J. Immunol. 138: 2826-2830. 2. Finkelman F, Morris S, Orekhova T, and Sehy D. 2003. The Cincinnati Cytokine Capture Assay (CCCA) for measurement of cytokine production in the mouse. In Current Protocols in Immunology. Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, and Strober W, eds. Greene Publishing Associates and Wiley-Interscience, New York, Unit 6.28, supplement 54. 3. Choo, S-W, and DH Conrad. 1997. A new multivalent B cell activation model - anti-IgD bound to FcgRI: properties and comparison with CD40L-mediated activation. International Immunol. 9: 239-248.