Anti α dystroglycan iih6c4

We performed open biopsy on the right deltoid in the patient. The muscle tissue was frozen and then cut at 7 μm sections. These sections were stained according to standard histological and enzyme histochemical procedures with hematoxylin and eosin (HE), modified Gomori Trichrome (MGT), periodic acidic Schiff (PAS), oil red O (ORO), nicotinamide adenine dinucleotide tetrazolium reductase, succinate dehydrogenease, cytochrome C oxidase, and esterase.
Sections were also immunostained with the following primary antibodies with standard procedures: dystrophin (DYS1: Rod domain; DYS2: C‐terminus; DYS3: N‐terminus, Novocastra, Newcastle‐upon‐Tyne, UK), dysferlin (NCLHamlet, Novocastra), MHC‐I (ab52922, abcam), anti‐α‐dystroglycan (IIH6C4, Millipore), anti‐GMPPB (HPA014657, Sigma). Additionally, anti‐β‐Spectrin (NCL‐SPEC1, Novocastra) was exploited as sarcolemma marker. A nondisease control was set and labeled at the same magnification and exposure.
Protein of muscle tissue lysate was extracted and the expression level of GMPPB was detected by western blotting with anti‐GMPPB.

Cryosections were stained with anti-dystrophin (ab15277, 1:100, Abcam), anti-BrdU-biotin conjugate (B35138, 1:100, Life Technologies), anti-laminin-α2 (4H8-2, 1:100, Sigma) and anti-α-dystroglycan (IIH6C4, 1:50, Millipore). Sections were fixed in ice-cold acetone for 10 min, washed in PBS-Tween (0.1% tween-20; Sigma-Aldrich), and blocked for 1 h in PBS supplemented with 20% goat serum (GeneTex), 0.1% tween-20 (Sigma-Aldrich), and 10 mg/ml BSA (Sigma-Aldrich). Primary antibodies were incubated overnight at 4°C in a humidified chamber. Sections were then washed and probed with the appropriate Alexa Fluor secondary antibody (Thermo Fisher) at a dilution of 1:500 for 1 h. Sections were mounted with Prolong Gold Mounting Media (Thermo Fisher) with 4^′,6-diamidino-2-phenylindole (DAPI) for nuclear staining. For BrdU immunostaining, sections were fixed in ice-cold acetone for 10 min, incubated in 2N HCl at 37°C for 30 min, and briefly neutralized with 0.15 M sodium tetraborate (Sigma-Aldrich); sections were blocked and incubated in the appropriate antibodies as described above and stained with a 10x DAPI stock prior to mounting in Prolong Gold Mounting Media.