Anti tdp 43 2e2 d3

Immunofluorescence labeling was performed on 6-μm-thick sections of paraffin-embedded temporal cortex samples from the brains. Before immunostaining, the sections were microwaved 2 × for 2 minutes each in 0.01 mol/L citrate buffer, pH 6.0, for antigen retrieval. Sections were incubated with anti-TDP-43 2E2-D3 (Abnova, Walnut) and anti-p65 antibodies (Santa Cruz Biotechnology, Santa Cruz) and subsequently with corresponding Alexa 488 and 633 antibodies (Molecular Probes, Eugene, OR). The nuclei were counterstained with Dapi (Invitrogen, Camarillo). Sections were observed by confocal laser microscopy (FV300 and FV1000, Olympus, Tokyo, Japan). Autofluorescence, such as lipofuscin pigments, were detected using 575–630 nm bandpass emission filter.

The following antibodies were used in this study: anti-actin (Millipore, Billerica, MA), anti-neuronal nuclear antigen (NeuN) (Chemicon International, Temecula, CA), anti-p65 (Santa Cruz Biotechnology, Santa Cruz, CA), anti-p65 (Invitrogen, Camarillo, CA), anti-phospho-p65 (phosphorylated at serine 536, Cell signaling technology, Danvers, MA), anti-tau 13 (Covance, Princeton, NJ), anti-tau CP-13 (phosphorylated at serine 202/threonine 205, gift from Dr. Peter Davies; Albert Einstein College of Medicine, Bronx, NY), anti-TDP-43 2E2-D3 (human-specific monoclonal antibody to total TDP-43; Abnova, Walnut, CA), and anti-C-TDP-43 (12892 polyclonal antibody to C-terminal TDP-43; ProteinTech Group, Chicago, IL).