Ultrathin‐layer isoelectric focusing of enzymes in liver samples of wagtails ( Motacilla flava, ssp. )

Wagtail samples ( Motacilla flava, ssp ; song bird) have been collected in different regios of central and southern Europe. Due to sampling extending over long periods of time it was necessary to investigate the effect of storing conditions (liquid nitrogen or ‐36 °C). Isozyme separation was done by polyacrylamide ultrathinlayer isoelectric focusing (100 μm). Liver homogenates were diluted in appropriate manner and 5 μl aliquots were used for separations. Gels for isoelectric focusing (T = 5.4, C = 2.8) containing different kinds of carrier ampholytec (Servalyt, Pharmalyte and Ampholines were cast on 12.5 × 20 cm pretreated glass plates according to the procedure of Radola [9]. Enzyme staining was performed in solution (ACP and esterases) or with an agarose overlay technique (LDH, ADH, MDH, PGM and GPI). Wagtail liver samples were chosen to optimize the electrophoretic conditions and to test the usefulness of different staining methods. This — and investigations presently under way on samples of pectoral muscle, heart, brain and blood — should allow us to elucidate genetic differences at the molecular level of the various subspecies of wagtails. The smaller total amount of protein available in some of these samples stresses the necessity of choosing a reproducible and sensitive, as well as rapid, electrophoretic technique.