This antibody gave a positive signal in the following human lysates:
HeLa Whole Cell, HeLa Nuclear, Jurkat Whole Cell, Jurkat Nuclear, HepG2 Whole Cell, HepG2 Nuclear, A431 Whole Cell and K562 Nuclear - tumor cell line.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 45 kDa).
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration. PubMed: 20736350
FunctionDNA and RNA-binding protein which regulates transcription and splicing. Involved in the regulation of CFTR splicing. It promotes CFTR exon 9 skipping by binding to the UG repeated motifs in the polymorphic region near the 3'-splice site of this exon. The resulting aberrant splicing is associated with pathological features typical of cystic fibrosis. May also be involved in microRNA biogenesis, apoptosis and cell division. Can repress HIV-1 transcription by binding to the HIV-1 long terminal repeat. Stabilizes the low molecular weight neurofilament (NFL) mRNA through a direct interaction with the 3' UTR.
Tissue specificityUbiquitously expressed. In particular, expression is high in pancreas, placenta, lung, genital tract and spleen.
Involvement in diseaseDefects in TARDBP are the cause of amyotrophic lateral sclerosis type 10 (ALS10) [MIM:612069]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology of ALS is likely to be multifactorial, involving both genetic and environmental factors. The disease is inherited in 5-10% of the cases.
DomainThe RRM domains can bind to both DNA and RNA.
Post-translational modificationsHyperphosphorylated in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU. Ubiquitinated in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU. Cleaved to generate C-terminal fragments in hippocampus, neocortex, and spinal cord from individuals affected with ALS and FTLDU.
Cellular localizationNucleus. In patients with frontotemporal lobar degeneration and amyotrophic lateral sclerosis, it is absent from the nucleus of affected neurons but it is the primary component of cytoplasmic ubiquitin-positive inclusion bodies.
Lane 1: Wild-type HAP1 cell lysate (40 µg) Lane 2: TDP43 knockout HAP1 cell lysate (40 µg) Lane 3: HeLa cell lysate (40 µg) Lane 4: Jurkat cell lysate (40 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab41881 observed at 48 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab41881 was shown to specifically react with TDP43 when TDP43 knockout samples were used. Wild-type and TDP43 knockout samples were subjected to SDS-PAGE. Ab41881 and ab8245 (loading control to GAPDH) were diluted at 1 µg/mL and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-TDP43 antibody (ab41881)Image from Shan X et al, Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16325-30. Epub 2010 Aug 24, Fig 2. DOI 10.1073/pnas.1003459107
ab41881 staining TARDBP in murine spinal cord by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissues fixed by 4% paraformaldehyde in phosphate buffer (pH 7.4) were cryoprotected by 30% sucrose in PBS (pH 7.4). Frozen sections (10 µm) of spinal cord tissues from three or more transgenic mice, respectively, were processed for immunofluorescence using ab41881. An Alexa Fluor 594 was used as secondary antibodies to visualize protein. Scale bar 20 µm. TARDBP is localized in the nucleus and forms intranuclear granular structures, arrowheads.
ICC/IF image of ab41881 stained human Hek293 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab41881, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Western blot - Anti-TDP43 antibody (ab41881)
All lanes : Anti-TDP43 antibody (ab41881) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : Jurkat nuclear extract lysate (ab14844) Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 6 : HepG2 nuclear extract lysate (ab14660) Lane 7 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 8 : K562 (Human) Nuclear Lysate - tumor cell line (ab29309)
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution