Purification procedure and monoclonal antibodies: two instruments for research on vertebrate porins.
On Western blots of skeletal muscle preparations of different vertebrate classes, four monoclonal anti-human type 1 porin antibodies recognize one single band of either 30.5 or 31 kDa, respectively. To confirm that it is eukaryotic porin which is labeled by the antibodies, we used a purification procedure developed for human type 1 porin for porins from skeletal muscle of shark, frog, and turkey. Applied to different mammalian species and tissues, this procedure exclusively provides type 1 porin. However, applied to shark skeletal muscle, it provides two porin isotypes in nearly equal amounts. In the case of frog skeletal muscle, the procedure provides mainly type 2 porin and a lower amount of type 1 porin. Applied to turkey skeletal muscle, the method provides exclusively type 2 porin. As demonstrated by two-dimensional Western blots, both shark and frog porin isotypes and the turkey type 2 porin are recognized by our antibodies. Furthermore, we elucidated the entire amino acid sequence of frog type 2 porin.
Reymann
S
S
Department of Immunochemistry, Department of Molecular Neuroendocrinology, Max Planck Institute for Experimental Medicine, Hermann-Rein-Strasse 3, Göttingen, D-37075, Germany. reymann@exmdi1.mpiem.gwdg.de
Kiafard
Z
Z
Rohm
B
B
Strutz
N
N
Hesse
D
D
Kratzin
H D
HD
Zimmermann
B
B
Thinnes
F P
FP
Hilschmann
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001001:Anura; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000737:chemistry; D018272:Porins / Q000302:isolation & purification*; D012754:Sharks; D014421:Turkey; D050994:Voltage-Dependent Anion Channels
D000911:Antibodies, Monoclonal; D018272:Porins; D050994:Voltage-Dependent Anion Channels