This work was funded by a grant to J.A.R. from the Jain Foundation Inc.
Improved immunoblotting methods provide critical insights into phenotypic differences between two murine dysferlinopathy models
Version of Record online: 14 JUL 2014
Copyright © 2014 Wiley Periodicals, Inc.
Muscle & Nerve
Volume 50, Issue 2, pages 286–289, August 2014
How to Cite
Mueller, A. L., Desmond, P. F., Hsia, R.-c. and Roche, J. A. (2014), Improved immunoblotting methods provide critical insights into phenotypic differences between two murine dysferlinopathy models. Muscle Nerve, 50: 286–289. doi: 10.1002/mus.24220
The authors declare no conflicts of interest.
- Issue online: 21 JUL 2014
- Version of Record online: 14 JUL 2014
- Accepted manuscript online: 23 FEB 2014 04:15AM EST
- Manuscript Accepted: 20 FEB 2014
Additional Supporting Information may be found in the online version of this article.
|mus24220-sup-0002-suppfig1.tif||2602K||Supporting Information Figure 1. Optimization of tissue homogenization for Immunoblotting dysferlin. (A) Homogenates of the right and left TA muscles of 2 A/WySnJ mice were prepared under several different conditions to identify conditions that give the most complete extraction of dysferlin with the least background noise. (B) The use of RIPA buffer and sonication result in the most complete extraction of dysferlin with the least amount of background noise.|
|mus24220-sup-0003-suppfig2.tif||3563K||Supporting Information Figure 2. Immunogold labeling of dysferlin is specific and places dysferlin at the ER, SR, Golgi networks. (A,B) Immunogold labeling performed with the Hamlet antibody gives clear and defined labeling of dysferlin and suggests that dysferlin is enriched at membranes close to the Z-disk, which have been reported to be part of the ER, SR, and Golgi networks. Labeling with non specific mouse IgG does not show any gold particles confirming that dysferlin labeling with Hamlet is specific. (C) Results from all pairwise comparisons for the statistical analyses shown in Figure 1I are provided.|
|mus24220-sup-0004-suppfig3.tif||7701K||Supporting Information Figure 3. Dysferlin is enriched in membranes close to the Z-disk. (A,B) The highest percentage of gold particles bound to the Hamlet antibody are present at the outer and inner Z-disk.|
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