Schedl Lab

Washington University Genetics

Glycine elution (affinity purification)

Elution off the coloumn with Glycine-Hcl (pH 2.3)

Adapted by Sudhir Nayak from Swathi Arur original.

Solutions

To make 0.1 M Glycine-HCl, pH 2.3:

Make 100mM Glycine, and pH with HCl to a pH of 2.3, alternatively use Glycine-HCl product.  Check pH before use.

Bind antibodies to your favorite protein column.

1) Wash column with 5 volumes TBS / PBS.

2) Wash column with 10 volumes Wash buffer / (500 mM NaCl, 20 mM Tris (pH 7.4), 0.1% Triton X-100).

3) Wash column with 5 volumes TBS  / PBS.

Elute bound antibodies.

1) Elute bound proteins with 2 mls glycine elution buffer.

2) Collect 10 x 200 ul fractions in tubes containing 50 ul 2 M Tris (pH 8.5).

3) Wash column with 10 volumes TBS / PBS.

4) Wash column with 5 volumes TBS / PBS + azide and store column at 4 degrees C.

  • Tim Schedl, Ph.D
    Department of Genetics
    Campus Box #8232
    Washington University School of Medicine
    4566 Scott Ave.
    St. Louis, MO 63110

    Contact

    Email: ts@genetics.wustl.edu
    PHONE: (314)362-6164 [lab]
    FAX: (314)362-7855