Description
IgM binds poorly to Protein A and, until recently, no readily available affinity chromatography product existed for one- step IgM purification. Standard methods for IgM purification generally are multi-step, tedious processes, or they are not effective for removing all of the major impurities present in IgM samples. • Pierce has developed an easy-to-use affinity matrix and buffer system to purify IgM. This matrix, an immobilized mannan binding protein (MBP) support, is most effective for purifying mouse IgM from ascites. Purified IgM can be obtained from a single pass over the affinity column. Human IgM will bind to the support and yield a product at least 88% pure by HPLC upon elution. The purification of IgMs from other species and mouse serum has not yet been optimized. Studies in Pierce labs show that MBP does not bind F(ab´)2 and Fab. • Mannan Binding Protein is immobilized on two supports: agarose and UltraLink Biosupport Medium. The Immobilized MBP is also available in a kit format – the Pierce IgM Purification Kit. The kit contains sufficient buffers to perform 10 purifications using a 5 ml column of Immobilized MBP. The Pierce IgM Purification Kit is easy-to-use and yields 90% pure mouse IgM (from ascites) with a very simple protocol. Immobilized MBP can be regenerated at least 10 times with no apparent loss of binding capacity. The figure below demonstrates the high purity of Immobilized MBP purified IgM from mouse ascites.
- Immobilized MBP Column 5 ml
- IgM Binding Buffer 800 ml
- IgM Elution Buffer 500 ml
- MBP Column Preparation Buffer 50 ml
- Column Extender