PSCP GA

Place 200 µl of 50% slurry protein A or protein G sepharose in the spin-X cups.

Spin at 13 000 rcf ~11 000 rpm 30’’. Discard the eluate.

Add 400 µl of protein A or protein G -specific binding buffer and wash for 5’, end-over end. Spin at 13 000 rcf ~11 000 rpm 30’’. Discard the eluate.

Repeat as above 2x more.

Dilute 200 µl of antibody (0,2 µg/µl) with 200 µl of protein A or protein G -specific binding buffer. Add to the protein A or protein G beads and incubate for 30’ end-over-end.

Spin at 13 000 rcf ~11 000 rpm 30’’ and discard the eluate.

Add 400 µl of protein A or protein G -specific binding buffer. Spin at 13 000 rcf ~11 000 rpm 30’’. Discard the eluate.

Repeat as above 2x more.

Add 400 µl of protein A or protein G -specific eluting buffer and wash for 5’, end-over end. Spin at 13 000 rcf ~11 000 rpm 30’’. Collect the eluate and add 40 µl of 1 M Tris-Cl, pH=8,8 to neutralize.

Repeat as above 2x more.

Measure the antibody concentration, concentrate and dialyse, or perform buffer exchange chromatography.