| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Technical Advance |
From the Neurobiology Department, University Medical School, Framlington Place, Newcastle upon Tyne, United Kingdom
A multiplex system of Western blotting is presented in which most
of the current muscular dystrophy proteins can be analyzed
simultaneously on one pair of blots. This represents a significant
improvement in efficiency and cost for this type of analysis. The final
diagnosis is more quickly achieved in patients where several possible
diagnoses are indicated after clinical appraisal, and those
with unusual presentations may be quickly resolved. The method uses a
biphasic polyacrylamide gel system, which enables the
corresponding blot to be probed simultaneously with a cocktail of
monoclonal antibodies. The gel is optimized so that large proteins of
more than 200 kd (eg, dystrophin, dysferlin,
and myosin heavy chain) can be analyzed in the top part, while
smaller proteins under 150 kd (eg, calpain 3, the 80-kd
fragment of laminin 
2 chain, all of the
sarcoglycans, and caveolin 3) are separated in the lower phase.
This basic system could be used for different combinations of
antibodies as new muscular dystrophy proteins are identified and
require examination. In addition, analysis of the laminin 2
chain of merosin showed that this protein was expressed as a doublet or
triplet set of bands in many patients with active muscle pathology.
This may indicate the existence of an embryonic isoform, which
is re-expressed in regenerating fibers.
This article has been cited by other articles:
 |
|
 |
|
  E. J. Groen, R. Charlton, R. Barresi, L. V. Anderson, M. Eagle, J. Hudson, M. S. Koref, V. Straub, and K. M. D. Bushby Analysis of the UK diagnostic strategy for limb girdle muscular dystrophy 2A Brain, December 1, 2007; 130(12): 3237 - 3249. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Illa, N. De Luna, R. Dominguez-Perles, R. Rojas-Garcia, C. Paradas, J. Palmer, C. Marquez, P. Gallano, and E. Gallardo Symptomatic dysferlin gene mutation carriers: Characterization of two cases Neurology, April 17, 2007; 68(16): 1284 - 1289. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Huang, S. H. Laval, A. van Remoortere, J. Baudier, C. Benaud, L. V. B. Anderson, V. Straub, A. Deelder, R. R. Frants, J. T. den Dunnen, et al. AHNAK, a novel component of the dysferlin protein complex, redistributes to the cytoplasm with dysferlin during skeletal muscle regeneration FASEB J, March 1, 2007; 21(3): 732 - 742. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. de Luna, E. Gallardo, M. Soriano, R. Dominguez-Perles, C. de la Torre, R. Rojas-Garcia, J. M. Garcia-Verdugo, and I. Illa Absence of Dysferlin Alters Myogenin Expression and Delays Human Muscle Differentiation "in Vitro" J. Biol. Chem., June 23, 2006; 281(25): 17092 - 17098. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. C. Brown, S. Torelli, M. Brockington, Y. Yuva, C. Jimenez, L. Feng, L. Anderson, I. Ugo, S. Kroger, K. Bushby, et al. Abnormalities in {alpha}-Dystroglycan Expression in MDC1C and LGMD2I Muscular Dystrophies Am. J. Pathol., February 1, 2004; 164(2): 727 - 737. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. T. Cooper, H. P. Lo, and K. N. North Single section Western blot: Improving the molecular diagnosis of the muscular dystrophies Neurology, July 8, 2003; 61(1): 93 - 97. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Poppe, L. Cree, J. Bourke, M. Eagle, L.V.B. Anderson, D. Birchall, M. Brockington, M. Buddles, M. Busby, F. Muntoni, et al. The phenotype of limb-girdle muscular dystrophy type 2I Neurology, April 22, 2003; 60(8): 1246 - 1251. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Aartsma-Rus, A. A.M. Janson, W. E. Kaman, M. Bremmer-Bout, J. T. den Dunnen, F. Baas, G.-J. B. van Ommen, and J. C.T. van Deutekom Therapeutic antisense-induced exon skipping in cultured muscle cells from six different DMD patients Hum. Mol. Genet., April 15, 2003; 12(8): 907 - 914. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. G. De Angelis, O. Sthandier, B. Berarducci, S. Toso, G. Galluzzi, E. Ricci, G. Cossu, and I. Bozzoni Chimeric snRNA molecules carrying antisense sequences against the splice junctions of exon 51 of the dystrophin pre-mRNA induce exon skipping and restoration of a dystrophin synthesis in Delta 48-50 DMD cells PNAS, July 9, 2002; 99(14): 9456 - 9461. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Yamada, F. Saito, H. Fukuta-Ohi, D. Zhong, A. Hase, K. Arai, A. Okuyama, R. Maekawa, T. Shimizu, and K. Matsumura Processing of {beta}-dystroglycan by matrix metalloproteinase disrupts the link between the extracellular matrix and cell membrane via the dystroglycan complex Hum. Mol. Genet., July 1, 2001; 10(15): 1563 - 1569. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Fanin, E. Pegoraro, C. Matsuda-Asada, R.H. Brown Jr., and C. Angelini Calpain-3 and dysferlin protein screening in patients with limb-girdle dystrophy and myopathy Neurology, March 13, 2001; 56(5): 660 - 665. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Torrente, J.-P Tremblay, F. Pisati, M. Belicchi, B. Rossi, M. Sironi, F. Fortunato, M. El Fahime, M. G. D'Angelo, N. J. Caron, et al. Intraarterial Injection of Muscle-derived CD34+Sca-1+ Stem Cells Restores Dystrophin in mdx Mice J. Cell Biol., January 22, 2001; 152(2): 335 - 348. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Dinçer, Z. Akçören, E. Demir, I. Richard, O. Sancak, G. Kale, E. Tan, J A. Urtizberea, and J. S Beckmann A cross section of autosomal recessive limb-girdle muscular dystrophies in 38 families J. Med. Genet., May 1, 2000; 37(5): 361 - 367. [Abstract] [Full Text]
|
|
|
|
|
|
K. M. D. Bushby The limb-girdle muscular dystrophies--multiple genes,multiple mechanisms Hum. Mol. Genet., September 1, 1999; 8(10): 1875 - 1882. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. D. Bushby Making sense of the limb-girdle muscular dystrophies Brain, August 1, 1999; 122(8): 1403 - 1420. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
