Cellulose Acetate Paper Research Articles

Metabolism of the developing retina: 7. Lactic dehydrogenase isoenzyme in the normal and degenerating retina a preliminary communication

The isoenzymes of retinal lactic dehydrogenase have been separated employing electrophoresis on cellulose acetate paper. The results confirm those of other workers using more elaborate procedures in that there are five fractions each of which is capable of reacting with either NAD or NADP. Greater activity is shown by all fractions with the former. Marked variations have been found in fraction five both during the development of the retina and between the retina of normal and retinitis pigmentosa rats. The significance of these findings is discussed with special reference to tissue differentiation.

PURIFICATION AND CHARACTERIZATION OF STAPHYLOCOAGULASE.

Zolli, Zeno, Jr. (Michigan State University, East Lansing), and Charles L. San Clemente. Purification and characterization of staphylocoagulase. J. Bacteriol. 86:527-535. 1963.-Separation and extreme purification of coagulase from Staphylococcus aureus strain 70 was achieved by using three cycles of dialysis in ethanol-water mixtures under controlled conditions, followed by molecular sieving through a column of Sephadex G-200. By manipulation of five variables (pH, ionic strength, temperature, protein, and ethanol concentration), the final preparation showed an approximate 3700-fold increase in activity per mg of protein. The successfully isolated coagulase containing 15.0% nitrogen was characterized serologically and chemically. By use of agar diffusion techniques, one zone of precipitation was obtained with the highly purified material. Additional confirmation of purity was evidenced by the appearance of a single peak with cellulose acetate paper electrophoresis with a barbital buffer at pH 8.6. Progressive and eventual elimination of carbohydrate, deoxyribonuclease, lipase, and phosphatase was observed through the four stages of purification. Temperature studies showed that the stability of each fraction was inversely related to its purity.

Microelectrophoresis using small volumes of buffer

The construction and use of a simple, economical micro electrophoresis apparatus is described in detail. With cellulose acetate paper as the supporting medium, electrophoresis requires 30 min. Dry batteries may be used as the power supply; at 14 V resolution is obtained in 5–7 h.

Electrophoretic Separation of Serum Protein Into Twelve Fractions

Recently it has been demonstrated that the electrophoretic separation of serum proteins can be improved. Thus an increase in the number of separated electrophoretic components, or zones, from the conventional six to nine was made possible by the introduction of a new buffer (TRIS-EDTA-boric acid), the TEB-buffer (Aronsson & Gronwall, 1957). We have now found that the number of separate zones obtained from a normal serum with this buffer on cellulose acetate strips (Oxoid) (Kohn, 1957) can be increased further through some minor modifications of the method. The concentration of the buffer has been reduced in relation to the buffer used in earlier experiments with cellulose acetate paper and the bulffer now has the fdlowitlg composition :