The members of the murine Cdx family (Cdx1, Cdx2, and Cdx4) are members of the caudal-type homeobox family of genes, which are homologues of the Drosophila 'caudal' gene required for anterior-posterior regional identity. The intestine-specific transcription factors Cdx1 and Cdx2 are candidate genes for directing intestinal development, differentiation, proliferation and maintenance of the intestinal phenotype. The relative expression of Cdx1 to Cdx2 protein may be important in the anterior to posterior patterning of the intestinal epithelium and in defining patterns of proliferation and differentiation along the crypt-villus axis. Expression of the Cdx1 homeobox gene in epithelial intestinal cells promotes cellular growth and differentiation. Cdx1 positively regulates its own expression. Cdx1 and Cdx2 are expressed in the small intestine and colon of fetus and adult. A decrease in human Cdx1 and/or Cdx2 expression is associated with colorectal tumorigenesis. Both Cdx1 and Cdx2 genes must be expressed to reduce tumorigenic potential, to increase sensitivity to apoptosis and to reduce cell migration, suggesting that the two genes control the normal phenotype by independent pathways. The human Cdx1 gene maps to chromosome 5q31-q33 and encodes a 265-amino acid protein.
Background References
1. Serra R W et al. A KRAS-directed transcriptional silencing pathway that mediates the CpG island methylator phenotype. Elife 3:E02313-E02313 (2014).
2. Yin Y et al. Impact of cytosine methylation on DNA binding specificities of human transcription factors. Science 356:0-0 (2017).
Western blot analysis of Cdx1 on Caco-2 (Human colorectal adenocarcinoma cell) cell lysate with Rabbit anti-Cdx1 antibody (ET7108-71) at 1/2,000 dilution.
Lysates/proteins at 15 µg/Lane. Exposure time: 3 minutes; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET7108-71, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 28 kDa Observed band size: 28 kDa
Immunocytochemistry analysis of LOVO cells labeling Cdx1 with Rabbit anti-Cdx1 antibody (ET7108-71) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Cdx1 antibody (ET7108-71) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Cdx1 antibody (ET7108-71) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-71) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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