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Preprint

Histological examination

FZ Francesco Zorzato
AR Alexis Ruiz

Last updated date: Jul 8, 2022 Views: 559 Forks: 0

original An abbreviated version of this protocol was published in eLife in Mar, 2022
Improvement of muscle strength in a mouse model for congenital myopathy treated with HDAC and DNA methyltransferase inhibitors
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1) Samples in PBS for 5min at room temperature (RT)
2) Block at RT for 30min (Blocking solution: 0.4%TritonX100 in PBS (containing 10 % goat serum))
3) Wash 3 x 5 min in PBS
4) Primary antibody 2h at RT (Primary antibody solution: 10 % Goat serum in PBS). 
5) wash 4 x 5 min
6) Secondary antibody 1h at RT (Secondary antibody solution: 10 % Goat serum in PBS)
7) wash 4 x 5min
8) add mounting medium (prolong gold antifade: Dako faramount aqueous mounting medium - ref S3025 )
9) put cover slip.

 

 

Primary Antibody mix 
Reagentdilution
BA-D5 - I1:100
SC-71 - IIA1:200
BF-F3 - IIB1:50
Laminin b1c1 (L9393)1:200

 

 

Secondary Antibody mix 
Reagentdilution
AF568 IgG1 GaM (21124)1:100
AF488 IgM GaM (21042)1:100
Dylight 405 IgG GaM (115-475-207)1:40
AF647 IgG DaRb (711-605-152)1:200

 

 

 

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Zorzato, F and Ruiz, A(2022). Histological examination. Bio-protocol Preprint. bio-protocol.org/prep1777.
  2. Ruiz, A., Benucci, S., Duthaler, U., Bachmann, C., Franchini, M., Noreen, F., Pietrangelo, L., Protasi, F., Treves, S. and Zorzato, F.(2022). Improvement of muscle strength in a mouse model for congenital myopathy treated with HDAC and DNA methyltransferase inhibitors. eLife. DOI: 10.7554/eLife.73718

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