Page "learned" Paragraph 143

Blood samples were allowed to clot at room temperature for 3 hr, centrifuged and the serum was removed.

The serum was measured volumetrically and subsequently dialyzed in the cold for at least 24 hr against three to four changes, approximately 750 ml each, of `` starting buffer ''.

This buffer, pH 8.6, was 0.005 M in Af and 0.039 M in tris(hydroxymethyl)-aminometha ( Tris ).

After dialysis the sample was centrifuged and the supernatant placed on a Af cm column of EEAE-cellulose equilibrated with starting buffer.

The DEAE-cellulose, containing 0.78 mEq of N/g, was prepared in our laboratory by the method of Peterson and Sober ( 7 ) from powdered cellulose, 100 - 230 mesh.

The small amount of insoluble material which precipitated during dialysis was suspended in approximately 5 ml of starting buffer, centrifuged, resuspended in 2.5 ml of isotonic saline and tested for antibody activity.