Sucrose Gradient Protocol for IgG/IgM separation? - Protocol for sucrose IgM separation (Jan/08/2005 )

Hi:

Does anybody have a protocol or a reference for the isolation of IgG and IgM from human sera using Sucrose Gradient Centrigugation (Yup, it's prehistorical but our system really, really needs it and the old Immunology manuals don't seem to go that far back)

Cheers,

RTRT

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If your still interested:

Immunol Commun. 1979;8(3):337-45. Related Articles, Links

Isolation of stable aggregates of IgG by zonal ultracentrifugation in sucrose gradients containing albumin.

Knutson DW, Kijlstra A, Lentz H, van Es LA.

Soluble heat aggregates of [125I]IgG (A-IgG) were prepared and separated by gel filtration or by zonal ultracentrifugation, and fractions containing different size aggregates were then analyzed by reultracentrifugation. A-IgG formed single narrow peaks with constant S rates when isolated, stored and analyzed in solution containing 0.5% serum albumin. Thus, stable homogeneous aggregates of IgG of known size can be simply prepared and should prove useful both as a model for immune complexes of specified sizes and as a standard for immune complex assays.

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