The alkaline phosphatase of leukocytes from normal subjects, patients with chronic granulocytic leukemia, and patients with reactive granulocytosis have been characterized by means of agarose column chromatography, sucrose density ultracentrifugation, starch gel electrophoresis, Michaelis constant for p-nitrophenyl phosphate, heat inactivation, inhibition by L-phenylalanine, Ouchterlony double ...

A purified and homogeneous preparation of rat AFP, as judged by both electrophoresis on Cellogel and immunoelectrophoresis, was separated into two components, AFPa and AFPb, by disc electrophoresis on 7% polyacrylamide gel. These two components had a definite difference in electrostatic net charge and gave only a single band on SDS-electrophoresis. Immunological reactivity or electrophoretic se...

A correlation has been observed between concanavalin A agglutination of various cell types and the presence of surface membrane galactosyltransferase (1-O-alpha-D-Galactosyl-myo-inositol:raffinose galactosyltransferase, EC 2.4.1.67) activity. Moreover, a reduction to less than 50% of cell surface galactosyltransferase activity occurred after treatment with concanavalin A; other cell surface gly...

Phytochrome was examined by immunochemical and spectroscopic techniques to detect differences between the protein moieties of red- and far red-absorbing phytochrome (P(r) and P(fr)). No differences in the reaction of P(r) and P(fr) with phytochrome antibody were discernible on Ouchterlony double diffusion plates. However, the microcomplement fixation assay showed a greater degree of antibody re...

A neutral endopeptidase (EC 3.4.24.5) that degrades alpha- and beta-crystallins occurs in mammalian lens. A procedure for purification of this enzyme from bovine lens is described. The enzyme appears to have a high molecular weight (Mr approximately equal to 700,000) and under denaturing conditions dissociates into at least eight polypeptide subunits with Mrs ranging from 24,000 to 32,000. A ne...

G. R. V. HUGHES plates. Sixteen wells 3 mm in diameter and 1-5 mm apart are cut in a row, and a trough 3 mm wide is cut parallel to the wells 3 mm from their cathodal side. Heat inactivated sera (8 .lI) are added to the wells, and a tissue extract containing soluble antigens (150 ,ul) is placed in the trough. We use human spleen extract4 or rabbit thymus extract at a final protein concentration...

The method for producing antigen-antibody precipitin bands in agar gel, due to Ouchterlony, is becoming increasingly useful in diagnostic immunology as well as in research into antibody-antigen reactions. For both these purposes a dimensionally accurate record of all bands, including faint lines, crossovers, and spurs, is necessary for correct interpretation of the plate. The dimensional accura...

The alkaline phosphatase of leukocytes from normal subjects, patients with chronic granulocytic leukemia, and patients with reactive granulocytosis have been characterized by means of agarose column chromatography, sucrose density ultracentrifugation, starch gel electrophoresis, Michaelis constant for p-nitrophenyl phosphate, heat inactivation, inhibition by L-phenylalanine, Ouchterlony double ...

The sera of patients who have received multiple transfusions may contain antibodies against normal human serum components (1, 2). The first example of such antibodies was reported in a patient (C. de B.) who had received approximately 50 transfusions for the treatment of a refractory anemia of unknown etiology (3). By means of the Ouchterlony double-diffusion technique, it was shown that the an...

DNA ligase has been purified to near-homogeneity from the extract of bovine thymus with a yield of 5%. The purified enzyme catalyzed the joining of single-stranded breaks in duplex DNA at a rate of 33 nmol of phosphodiester bonds/min/mg of protein. The purified enzyme was homogeneous as judged by polyacrylamide gel electrophoresis and Ouchterlony double diffusion analysis. The enzyme is compose...