Anti tdp 43 antibody 10782 2 ap

Mouse tissues (brain, spinal cord, liver) were dissected, snap-frozen and homogenized in SDS/Urea Buffer (0.5% SDS/8M Urea in Phosphate Buffered Saline) for TDP-43 immunodetection or RIPA Buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% NP-40, 0.25% Na deoxycholate, 1 mM EDTA, 0.1% SDS, Protease Inhibitor Cocktail P8340 from Sigma diluted 1∶50) for Masp-2 immunodetection. Homogenates were analyzed on 10% (w/v) SDS-polyacrylamide gels and transferred to PVDF membranes which were then blocked for 1 hour at room temperature with 5% (w/v) skim milk powder in Tris-buffered saline (TBS). The membranes were then incubated overnight at 4°C with primary antibodies diluted in blocking solution as follows: mouse anti-mannan-binding Lectin (MBL) monoclonal antibody (MAB 3808) at 1∶5000 dilution; anti-TDP-43 antibody (10782-2-AP, Proteintech) diluted 1∶2000; or anti-Actin clone C4 (MAB 1501) diluted 1∶5000. Immunoblots were then washed with TBS containing 0.05% Tween 20 and incubated for 1 hour at room temperature with anti-mouse or anti-rabbit IgG conjugated to peroxidase diluted 1∶5000 in blocking solution. Antibody binding was detected with Western Lightning Plus ECL detection system (Perkin Elmer).