PMID:
8740185
Authors:
Lutomski D, Caron M, Cornillot JD, Bourin P, Dupuy C, Pontet M,
Bladier D, Joubert-Caron R.
Title:
Identification of different galectins by immunoblotting after two-dimensional
polyacrylamide get electrophoresis with immobilized pH gradients.
Journal:
Electrophoresis. 1996 Mar;17(3):600-6. doi: 10.1002/elps.1150170331.
Abstract:
Vertebrate soluble beta-galactoside-binding lectins form a growing protein family
that recently have been named galectins. Seven different galectins have been
sequenced and characterized in mammals, and there is compelling evidence for the
existence of other members of this lectin family. Three among six galectins are
homodimers with (i) an identical subunit of a relative molecular mass of about
14500, and (ii) amino acid sequence homologies giving rise to possible
immunochemical cross-reactivities. They are indistinguishable from each other by
conventional sodium dodecyl sulfate-polyacrylamide gel electrophoresis
(SDS-PAGE), even when followed by immunoblotting. However, their different
isoelectric points allow their identification using isoelectric focusing and
two-dimensional (2-D) polyacrylamide gel electrophoresis. A strategy was
developed to identify these galectins in crude extracts from cells and tissues,
based on the two-dimensional electrophoresis with immobilized pH gradient
(IPG-Dalt) analysis of the specific spots of purified galectins and of the spots
of crude extracts, after silver staining. In addition, 2-D immunoblotting using
anti-galectin 1 (Gal-1) and anti carbohydrate-binding protein 15 (CPB15)
antibodies were performed on brain and leukemia cells (HL60) allowing an
identification of related polypeptides. Our results indicate that the use of
IPG-Dalt provides a suitable reproducibility and allows the detection of
galectins or other galactoside-binding proteins even at basic pIs.
This page is constructed based on the NCBI service.
