Recombinant Anti-Tbx3 Antibody [RP23040141] (ab313456)

Enrichment of endogenous TBX3 protein at specific gene loci using ab313456: Chromatin Immunoprecipitation (ChIP) was performed using ab313456(5 ug) on sheared chromatin from 2 million HEPG2 cells. Normal Rabbit IgG (1 ug) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system with optimized PCR primer pairs for the different loci of the active P21 promoter region used as positive control target genes, and the region of the inactive MYOD, SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

For immunofluorescence analysis, MCF7 cells were fixed and permeabilized for detection of endogenous TBX3 using AB313456 (2 ug/ml) and labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (1:2000). Panel a) shows representative cells that were stained for detection and localization of TBX3 protein (green), Panel b) is stained for nuclei (blue) using SlowFade DAPI. Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of TBX3. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.