Rabbit anti tdp 43 10782 2 ap

The following antibodies were used for immunoblotting: mouse anti-TurboGFP (TA150041) [Origene]; rabbit anti-TDP-43 (10782-2-AP) [Proteintech]; mouse anti-Lamin A/C (05-714) [EMD Millipore]; rabbit anti-beta Actin (ab16039) [Abcam]; rabbit anti-NF-κB p65 (Ser536) (3033), rabbit anti-NF-κB (8242) and HRP-conjugated anti-α-tubulin (9099) [Cell Signaling Technology]; and rabbit anti-PTP1B (sc-14021), HRP-conjugated anti-rabbit IgG (sc-2004) and HRP-conjugated anti-mouse IgG (sc-2005) [Santa Cruz]. The following antibodies were used for immunocytochemistry (ICC): rabbit anti-MAP2 (1:500; AB5622) and TRITC-conjugated phalloidin (1:500; 90228) [Millipore]. The following antibody was used for immunohistochemistry (IHC): FITC-conjugated anti-HRP (1:150; 123-095-021) [Jackson ImmunoResearch Laboratories]. The following antibodies were used for neutralizing target proteins: rabbit anti-IL-1β (ab9722) [Abcam]; rat anti-TNF-α (14-7321-81) [Invitrogen]; rabbit anti-IgG (2729) and mouse anti-IgG (5415) [Cell Signaling Technology]; and mouse anti-IL-6 (sc-57315) and rat anti-IgG (sc-2026) [Santa Cruz].

HEK293T (#CRL-3216) cells were acquired from ATCC, was authenticated and tested negative for mycoplasma contamination. H293T cells were cultured on coverslips coated with poly-L-lysine (Sigma-Aldrich). Cells were transfected with either mRuby, NUP62-mRuby, HA-eGFP, Nup54-HA-eGFP, pEGFP-SF2/SRSF1 (Addgene, #17990), FLAG-NM23-H1/NME1 (Addgene, #25000), Frt-V5-HspB2 (Addgene, #63103), or mRFP-FKBP1A (Addgene, #67514) expression plasmids using TurboFect (Thermo Fisher Scientific), following the manufacturer’s instructions. Cells were washed and incubated with primary antibodies overnight: rabbit anti-TDP-43 (10782–2-AP), 1:1000 (Proteintech); or rat anti-phospho TDP-43 (MABN14), 1:1000 (Millipore, Sigma-Aldrich). Cells were washed and incubated with secondary antibodies: anti-rabbit Alexa Flour 647, 1:100 (Life Technologies, #1660844); anti-rat Alexa Flour 568 (Invitrogen, Cat # A-11077). Coverslips were mounted onto slides using Fluoroshield with DAPI (Sigma-Aldrich, #F6057). Cells were imaged with a NIKON A1 eclipse T_i confocal microscope.