This is a knockout-validated antibody summary, based on the publication "Rbfox Splicing Factors Promote Neuronal Maturation and Axon Initial Segment Assembly", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG1

Company: EMD Millipore

Antibody: NeuN

Catalog number: MAB377

Summary: Mouse monoclonal IgG1 against purified cell nuclei from mouse brain. Reacts with mouse, rat, human, avian, chicken, ferret, pig, and salamander. Suitable for western blot, immunocytochemistry/immunofluorescence, immunohistochemistry, immunohistochemistry (paraffin), immunoprecipitation and flow cytometry.

Western blot | Immunocytochemistry

ICC: paraformaldehyde fixation (4% paraformaldehyde in 1x PBS) for 10 min at room temperature.

ICC: 1x PBS, 10% horse serum, 0.2% Triton X-100, 0.05% sodium azide for 30 minutes.

ICC: antibodies were diluted in antibody buffer (1x PBS, 5% horse serum, 0.2% Triton X-100, 0.05% sodium azide) and applied overnight at 4°C in humidified chamber.

ICC: secondary antibodies diluted in antibody buffer for 2 hours at 4°C.

Laser-scanning confocal microscope (LSM 800, Zeiss) using a 40x oil-immersion objective.

Depletion of Rbfox proteins was confirmed by immunoblot analysis in in vitro differentiated neurons; GAPDH is a loading control. Validation of Rbfox1-3 protein depletion in Rbfox tKO motor neurons based on immunostaining analysis on day 5. Neuronal morphology was visualized by expression of GFP from transfected plasmid and motor neurons were identified by antibody staining of the endogenous Hb9 protein. Depletion of Rbfox proteins is determined by antibody that specifically recognizes each family member. Please see Figure 2a and S1d in the article [1].