Flow Cytometry - Anti-Ctip2 antibody [25B6] (ab18465)

Overlay histogram showing Jurkat cells stained with ab18465 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18465, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG (2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

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Immunocytochemistry/ Immunofluorescence - Anti-Ctip2 antibody [25B6] (ab18465)

Neonatal mouse hippocampal neurons stained with ab18465 (top panel - green) and Ctip1 antibody (middle - red). Bottom panel is overlay of ab18465 and Ctip1 antibody staining - yellow indicates co-localisation, green is ab18465 alone and red is Ctip1 antibody alone.

Western blot - Anti-Ctip2 antibody [25B6] (ab18465)

Western blot using ab18465 on nuclear extract from Jurkat cells immunoprecipitated with anti-Sir2 antibody.

Two bands are seen which may correspond to two CTIP2 transcripts present in Jurkat cells as previously reported (Bernard et al. 2001).

Western blot using ab18465 on nuclear extract from Jurkat cells immunoprecipitated with anti-Sir2 antibody.

Two bands are seen which may correspond to two CTIP2 transcripts present in Jurkat cells as previously reported (Bernard et al. 2001).

All lanes:

Western blot - Anti-Ctip2 antibody [25B6] (ab18465)

Predicted band size: 96 kDa

Immunocytochemistry/ Immunofluorescence - Anti-Ctip2 antibody [25B6] (ab18465)

Neonatal Mouse Hippocampal Neurons (Harvested at P1, grown 5d in culture on glial cell feeder layer).

Red is beta tubulin staining.

Green is ab18465.

Immunocytochemistry/ Immunofluorescence - Anti-Ctip2 antibody [25B6] (ab18465)

Immunofluorescence staining of Ctip2 using ab18465 in Jurkat cells (+ve expression control, top panel) and Daudi cells (-ve expression control, bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton-X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18465 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150084) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).