UNIT 2.9 Isolation of Murine and Human Immunoglobulin M and Murine Immunoglobulin D
Published Online: 1 APR 2009
Copyright © 2009 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Andrew, S. M., Titus, J. A., Amin, A. and Coico, R. 2009. Isolation of Murine and Human Immunoglobulin M and Murine Immunoglobulin D. Current Protocols in Immunology. 85:III:2.9:2.9.1–2.9.8.
- Published Online: 1 APR 2009
- Published Print: APR 2009
This unit describes two classical protocols for the purification of IgM—dialysis of ascites fluid, tissue culture medium, or bioreactor supernatants against distilled water to precipitate pure IgM, and ammonium sulfate precipitation. Both protocols can be followed by size-exclusion chromatography to obtain a highly purified product. Recently, an affinity method for purification of IgM has been developed using mannan-binding protein, and is described here. The third approach presented is a one-step IgD purification method, designed specifically for murine derived samples, that uses Sepharose coupled to lectin derived from the seeds of Griffonia simplicifolia-1. This represents a simple, rapid, and gentle, approach to isolating this highly labile immunoglobulin from IgD-containing ascites or hybridoma sources. Curr. Protoc. Immunol. 85:2.9.1-2.9.8. © 2009 by John Wiley & Sons, Inc.