This is a knockout-validated antibody summary, based on the publication "RNA m6A methylation regulates the ultraviolet-induced DNA damage response", as cited below [1]. This antibody was also validated for Western blot in the article "Ubiquitination Regulates the Proteasomal Degradation and Nuclear Translocation of the Fat Mass and Obesity-Associated (FTO) Protein" (see fgure 3b) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).
Rabbit monoclonal IgG
Company: Abcam
Antibody: FTO
Catalog number: ab126605
Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to human FTO (N terminal). Reacts with human by western blot, immunohistochemistry (paraffin), immunocytochemistry/immunofluorescence. Unsuitable for flow cytometry.
Western blot | Immunocytochemistry
ICC: cells were fixed with PBS containing 4% paraformaldehyde (PFA) for 10 min at room temperature.
WB: 5% skim milk in TBS-T (Tris-Buffered Saline (TBS), 0.05% Tween-20) overnight.
ICC: PBS containing 1% BSA, 10% FBS, 0.25% Triton-X100, 0.02% NaN3.
ICC: 3 μg/ml in blocking buffer at room temperature for 1–2 h or at 4°C overnight.
WB: 1/5,000 dilution appropriate peroxidase-conjugated anti-rabbit secondary antibody (EMD Millipore) for 1 hour.
Fluorescent secondary antibodies (conjugated with Alexa Fluor 350, 488, 594, or 647; Invitrogen, cat. #s A-21244, A-11037, A11032, A11034, A11029).
WB: Western Lightning Plus ECL reagents (PerkinElmer) according to manufacturer’s instructions, and exposed to film.
ICC: Images were captured on Nikon Eclipse Ti microscope.
a, WT or FTO KO U2OS cells were subjected to UVA laser microirradiation, incubated at 37°C for 4 min, then stained for FTO and γH2A.X as indicated. Middle panel shows a representative γH2A.X-positive cell with FTO colocalizing at the damage site (arrow). The percentage of cells showing colocalization of signals is indicated on the right. b, Western blot of FTO in WT or FTO KO U2OS cells. Actin is shown as a loading control. Please see Extended Data Figure 3 in the article [1].
αMETTL3 (ABclonal, A8370), 0.5 μg/ml was KO validated, western blot. Extended Data Figure 2
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