I do not have access to chromatography. I have read that zinc or copper will precipitate the albumin.
You can use G-protein coupled to Sepharose beads. It is an easy and fast, just mix the suspension of protein G-Sepharose with your material then centrifuge and wash. The product label (protein G-Sepharose describes the steps). As you may know the protein G has affinity to IgG that is the most abundant Immunoglobulin in serum.
We used ti separate immunoglobulin from serum this way, with:
The ammonium sulphate globulin precipitation technique. -Manipulations are carried out at room temperature. Equal volumes of mouse serum, usually 50 ul, made up to a volume of 0.1 ml with 0*1 M phosphate buffered saline pH 7.3 in conical tubes used (were of 4 ml capacity, Gelman Hawksley Ltd). After incubation, 0-2 ml saturated (NH4)2SO4 was added to each tube, centrifuging at 15,000 rev/min for 5 minutes in a Beckman 152 Microfuge. The supernatant containing albumin is removed and the precipitate, now immunoglobulin enriched, is washed twice with half-saturated (NH4)2SO4.
Reference
Immunology, 1972, 22, 747.
The use of Ammonium Sulphate Globulin Precipitation
for Determination of Affinity of Anti-Protein Antibodies
in Mouse Serum
M. W. STEWARD AND R. E. PETTY
Thanks for the input, we have eliminated about 95% of any no immunoglobulin fractions.
Pierce has a kit that utilizes the Cibacron Blue method.
Pierce Blue Albumin Removal Kit
The Same advice to you Jeff:
We used to separate immunoglobulin from serum albumin this way:
The ammonium sulphate globulin precipitation technique. -Manipulations are carried out at room temperature. Equal volumes of mouse serum, usually 50 ul, made up to a volume of 0.1 ml with 0*1 M phosphate buffered saline pH 7.3 in conical tubes used (were of 4 ml capacity, Gelman Hawksley Ltd). After incubation, 0-2 ml saturated (NH4)2SO4 was added to each tube, centrifuging at 15,000 rev/min for 5 minutes in a Beckman 152 Microfuge. The supernatant containing albumin is removed and the precipitate, now immunoglobulin enriched, is washed twice with half-saturated (NH4)2SO4.
Reference
Immunology, 1972, 22, 747.
The use of Ammonium Sulphate Globulin Precipitation
for Determination of Affinity of Anti-Protein Antibodies
in Mouse Serum
M. W. STEWARD AND R. E. PETTY
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