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 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).

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

Red is beta tubulin staining.
Green is 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG (2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

All lanes : Anti-Ctip2 antibody [25B6] (ab18465) at 1/500 dilution

Lanes 1-2 : Mouse brain tissue lysate at 1.5 µg
Lane 3 : Mouse brain tissue lysate at 3 µg

Secondary
All lanes : IRDYE 680-conjugated Donkey Anti-Rat polyclonal. at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 95 kDa
Observed band size: 100,110 kDa why is the actual band size different from the predicted?


Exposure time: 10 minutes

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