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Brown, J. C., Spragg, J. H., Wheeler, G. N. 
ORCID: https://orcid.org/0000-0002-4335-8577 and Taylor, P. W.
(1990)
Identification of the B1 and B2 subunits of human placental laminin and rat parietal-yolk-sac laminin using antisera specific for murine laminin-beta-galactosidase fusion proteins.
Biochemical Journal, 270 (2).
pp. 463-468.
ISSN 0264-6021
Abstract
Antisera raised against fusion proteins consisting of murine laminin B1 and B2 subunit sequences fused to the C-terminus of Escherichia coli beta-galactosidase were tested for their subunit specificity on Western blots of deglycosylated murine Engelbreth-Holm-Swarm (EHS) laminin. The antisera raised against B2 subunit sequences (anti-XLB2.1 and anti-XLB2.2) bound only to the EHS laminin B2 subunit. One of the antisera raised against B1 subunit sequences (anti-XLB1.2) was specific for the B1 subunit, whereas two others (anti-XLB1.1 and anti-XLB1.3) cross-reacted with the EHS laminin B2 subunit. Gold-labelled heparin-albumin was shown to bind specifically to the A subunit of deglycosylated EHS laminin on Western blots. These reagents were used to identify the homologous subunits in rat parietal-yolk-sac laminin and human placental laminin. The anti-(fusion protein) antisera identified the B1 and B2 subunits of the rat laminin, and these were similar in size to the murine EHS B subunits. Human placental laminin gave bands of 400, 340, 230, 190 and 180 kDa on reducing SDS/PAGE. The anti-(fusion protein) antisera identified the 230 and 190 kDa bands as the B1 and B2 subunits respectively. Gold-labelled heparin-albumin bound to the 400, 340 and 190 kDa bands of human placental laminin and so did not unambiguously identify a single A subunit. The human placental laminin may contain a mixture of isoforms, with alternative subunits substituting for the A subunit.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | amino acid sequence,animals,antibody specificity,western blotting,polyacrylamide gel electrophoresis ,escherichia coli,galactosidases,glycosylation,humans,immune sera,laminin,mice,molecular sequence data,molecular weight,placenta,rats,recombinant fusion proteins,nucleic acid sequence homology,yolk sac,beta-galactosidase |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Cells and Tissues |
| Depositing User: | Pure Connector |
| Date Deposited: | 11 Aug 2015 16:26 |
| Last Modified: | 19 Apr 2023 00:48 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/54072 |
| DOI: | 10.1042/bj2700463 |
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