Elution off the coloumn with Glycine-Hcl (pH 2.3)
Adapted by Sudhir Nayak from Swathi Arur original.
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.