IgM Purification kit #C8056-M
IgM Purification Kit (Cat# C8056-M; ready-to-use solution; store at 4 °C) Introduction : IgM antibody is prominent in early immune responses to most antigens and predominates in
certain antibody responses such as 'natural' blood group antibodies. IgM is the major
immunoglobulin expressed on the surface of B cells. IgM normally constitutes about 10% of
serum immunoglobulins.
ABSbioTM ready-to-use IgM Purification Kit is formulated for rapid, purification of high-quality IgM
from plasma, serum, ascites fluid and tissue culture media, with no binding to other
immunoglobulins. The supplied buffers system provide for maximum immunoglobulin binding and
elution efficiency with the IgM affinity matrix resin column. IgM affinity matrix exhibits high affinity
binding to the Fc part of IgM. The high selectivity and yields obtained using IgM Affinity Matrix
enable a robust, fast, and efficient purification process in one step from many species (including
human, mouse and rat), and the purified IgM can be used for antibody label, immunoprecipitation
assays and immunotherapy. Kit Components IgM Affinity Matrix Column: 1 mL Binding Buffer: 200 mL Neutralization Buffer: 10 mL Elution Buffer: 100 mL Storage and Handling: Store all of the components at 4 °C. Shelf Life: 12 months after receipt. Specifications Matrix: Cross-linked poly(styrene-divinylbenzene) Bead size range: 50 um Ligand: IgM affinity ligand, free of animal components Binding capacity: ~ 6 mg/mL (IgM from serum) Reusable: stable ligand and resins can be regenerated and reused at least 10 times without
significant decline in binding capacity SDS-PAGE of purified Human IgM Maximum column capacity per purificationspecies mg of IgMHuman6.0-12Rat6.0-12Mouse6.0-12Applications :Purification of monoclonal and polyclonal antibodies; Immunoglobulin Affinity Purification,
ImmunoprecipitationProtocol Preparation of Antibody Samples Average IgM concentration in human serum is about 1.5 mg/mL. To ensure proper ionic strength
and pH for optimal binding, mix ascites fluid, serum, or tissue culture supernatant sample with
Binding Buffer at a ratio of 1:3. Centrifuge samples at 10,000 x g for 5 minutes to remove any
insoluble material and use only the clear supernatant for antibody purification. 1.The resin can be used at 2-25 ºC. Pour about 1 mL IgM affinity matrix into the column. Remove
the bottom cap to allow the storage solution to drain through the column.2. Slowly add 5-10 mL of Binding Buffer to the top of the resin. Allow the column to drain until
liquid level drops to resin level. 3. Add a clarified, diluted antibody sample (5-10 mL) to the column and allow it to slowly pass
through the immobilized ligand (Maximum column capacity per purification is about 6-12 mg IgM). 4. Add 10 mL of Binding Buffer and allow it to pass through the column to wash away nonbound
components (or until you see a stable baseline). 5. Add 4 mL of Elution Buffer and collect 1 mL elution fractions. Immediately add 100 µL of
Neutralization Buffer to each of the 1 mL elution fractions. 6. Measuring OD 280nm or Protein assay to identify and combine elution fractions that contain
the purified antibody. (Optional) Alternatively, use a desalting column or dialysis to exchange the
purified antibody into a more suitable buffer for long-term storage. 7. Regenerate the column by washing with 10 mL of Elution Buffer and followed by 5 mL of 20%
Ethanol. When approximately 1 mL of solution remains above the resin, cap the column and store
upright at 4ºC. Columns may be regenerated a minimal of 9 times without significant loss of
binding capacity.