Phosphorylation-dependent TDP-43 antibody detects intraneuronal dot-like structures showing morphological characters of granulovacuolar degeneration

Comparison of phosphorylated TDP-43-positive inclusions in oculomotor neurons in patients with non-ALS and ALS disorders

“…Samples were fixed with 4% paraformaldehyde in phosphate-buffered solution (PBS) (pH 7.4) and embedded in paraffin. Five-μm-thick transverse paraffin sections were prepared for Hematoxylin-Eosin (H&E) staining and then immunohistochemistry, which was carried out using a rabbit polyclonal anti-phosphorylationdependent TDP-43 (pTDP-43) antibody (1:3000) generated in our laboratory [23]. For enhancement, the samples were autoclaved for 5 min before reaction with the antibody.…”

“…These findings suggest that abnormal phosphorylation of TDP-43 should be a critical step in the pathogenesis of ALS. We have recently generated an antibody recognizing phosphorylated TDP-43 (pTDP-43) at position 409 and 410 [23]. Our antibody detects a single band at about 45 kDa and smaller fragments at about 25 kDa with smears in ALS and AD samples, and demonstrates dot-like inclusions in the brains of AD patients and aged subjects [23].…”

Involvement of endoplasmic reticulum stress defined by activated unfolded protein response in multiple system atrophy

“…The commercially available antibodies used in this study were anti-phosphorylated pancreatic ER kinase (pPERK) (phosphorylated PERK at threonine 981) (rabbit polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA) [6,7], anti-phosphorylated eukaryotic initiation factor 2α (eIF2α) (phosphorylated at serine 51, rabbit polyclonal; Cell Signaling, Danvers, MA) [23], anti-phosphorylated inositol requiring enzyme 1α (IRE1α) (phosphorylated at serine 724, rabbit polyclonal; Novus Biologicals, Littleton, CO) [7], anti-tubulin polymerizationpromoting protein (TPPP/p25α) (rabbit monoclonal; Epitomics Inc., Burlingame, CA) [24], anti-transferrin (rabbit polyclonal; Dako Cytomation, Glostrup, Denmark) [25,26], anti-Olig2 (rabbit polyclonal; IBL, Takasaki, Japan) [27], anti-glial fibrillary acidic protein (GFAP) (rabbit polyclonal;Dako Cytomation, Glostrup, Denmark) [28], anti-α-synuclein (mouse monoclonal; Invitrogen, Carlsbad, CA) [29], antiubiquitin (rabbit polyclonal; Dako Cytomation, Glostrup, Denmark) [30,31], anti-tau phosphorylated (AT8) (mouse monoclonal; Innogenetics, Ghent, Belgium) [32,33], anti-phosphorylated glycogen synthase kinase (pGSK3) (mouse monoclonal; Upstate Biotechnology, Lake Placid, NY) [33,34], anti-pSmad2/3 (phosphorylated at serine 423 and 425) (goat polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA) [33,35], and anti-phosphorylated neurofilament (SMI-31) (mouse monoclonal; Sternberger Monoclonals, Baltimore, MD) [33,36]. The generation and characterization of the antibody that recognizes phosphorylated TAR-DNA-binding protein 43 (pTDP-43) at positions 409 and 410 (A2) was described elsewhere [33].…”

A Japanese ALS6 family with mutation R521C in the FUS/TLS gene: A clinical, pathological and genetic report

“…However, no pTDP-43-positive inclusions, such as skein-like or round inclusions, were observed in the cytoplasm of anterior horn cells. Antibodies used were a rabbit polyconal antibody against FUS (A300-302A, 1:250, Bethyl, Montgomery, TX), a rabbit polyclonal antiubiquitin antibody (1:2,000, Dako, Denmark), a guinea pig monoclonal antibody against the amino-terminus of p62 (1:2000, Progen Biotechnik, Heidelberg, Germany) and a rabbit polyclonal antiphosphorylated TDP-43 (pTDP-43) antibody [22].…”