Purification procedure and monoclonal antibodies: Two instruments for research on vertebrate porins

Citation
S. Reymann et al., Purification procedure and monoclonal antibodies: Two instruments for research on vertebrate porins, ANALYT BIOC, 274(2), 1999, pp. 289-295
Citations number
30
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
ANALYTICAL BIOCHEMISTRY
ISSN journal
0003-2697 → ACNP
Volume
274
Issue
2
Year of publication
1999
Pages
289 - 295
Database
ISI
SICI code
0003-2697(19991015)274:2<289:PPAMAT>2.0.ZU;2-P
Abstract
On Western blots of skeletal muscle preparations of different vertebrate cl asses, four monoclonal anti-human type 1 porin antibodies recognize one sin gle band of either 30.5 or 31 kDa, respectively. To confirm that it is euka ryotic porin which is labeled by the antibodies, we used a purification pro cedure 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 sha rk 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-dime nsional Western blots, both shark and frog porin isotypes and the turkey ty pe 2 porin are recognized by our antibodies. Furthermore, we elucidated the entire mine acid sequence of frog type 2 porin. (C) 1999 Academic Press.