Antibodies to small peptide/haptens are generally raised by coupling to a large carrier proteins such as KLH. Antibodies are produced to both KLH and the peptide/hapten. Anti-KLH may give non-specific signals in various immunoassays. Anti-KLH antibodies can be removed by solid phase immunoaffinity column chromatography over the KLH-Agarose.
Binding Capacity:Process 2-5ml of high titer antiserum or 10mg IgG over 1ml of K0300-05 The unbound fraction may be re-processed if necessary. The column can be used many times following standard binding, elution, and regeneration conditions.
Recommended Usage of the Affinity Column:K0300-05 is supplied in 2ml plastic column that is capped at the bottom. Break off the flat tip to allow the buffer to drain. Allow sample to flow through column. A yellow cap is also provided to cap the clumn whe not in use or to stop the column. It is also possible to cap the column with parafilm if the yellow cap is lost.
Removal of KLH Antibodies:Note: Purification may be performed at 4°C or RT
1. Dilute 2-5ml antiserum with PBS at 1:2
2. Apply diluted antibody to column
3. Recycle Flow-Through 2-3 times
4. Anti-KLH will bind to the column
5. The antibody of interest is in the Flow Through
6. Compare the Flow Through antibody with unprocessed antiserum to confirm the removal of anti-KLH antibodies
If necessary, recycle the column to complete the removal of anti-KLH antibodies.
Regeneration of Column:1. Apply 10-20ml of Elution Buffer (0.1M Gly-HCl, pH 2.5
