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Fred Slape after brain surgery

Retired Sgt. 1st Class Frederick T. Slape shows off the scar on his head after having a tumor removed from his brain on Sept. 25, 2015.

http://game-rapidshare.com/Gagliardi-grandpa-of-Todesco-from-Pier?Alkemagra=116

Western blotting

Heart and cell homogenates (30 ?g protein), or isolated cytosolic (20 ?g protein), mitochondrial (15 ?g protein), and nuclear (20 ?g protein) fractions were electrophoresed on a 4–15% Mini-Protein TGX gel using a Mini-PROTEAN electrophoresis chamber (Bio-Rad), and proteins were transferred to a PVDF membrane using a Criterion Trans-blot System (Bio-Rad). Membranes were blocked with 5% non-fat dry milk (NFDM) in 50 mM Tris-HCl (pH 7.5) / 150 mM NaCl (TBS), washed with TBS-0.1% Tween 20 (TBST) and TBS, and incubated overnight at 4°C with antibody clones listed in Table C of S1 Table. Antibodies from Santa Cruz were used at 1:200 dilutions, and all other antibodies were used at 1:1000 dilution in TBST-5% NFDM. Membranes were washed with TBST and TBS, incubated with HRP-conjugated secondary antibody (1: 10,000 dilution, Southern Biotech, Birmingham AL), and images were acquired by using an Image Quant LAS4000 system (GE Healthcare, Pittsburgh MA). Immunoblots were subjected to Ponceau S staining to confirm equal loading and transferring of samples. Densitometry analysis of protein bands was performed using a Fluorchem HD2 Imaging System (Alpha Innotech, San Jose CA), and normalized against GAPDH (tissue homogenates and cytosolic fractions), COIV (mitochondrial fractions) or Lamin B (nuclear fractions).

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