General description

Homeobox protein CDX-1 (UniProt P47902; also known as Caudal-type homeobox protein 1) is encoded by the CDX1 gene (Gene ID 1044) in human. CDX1 is a gut transcription factor crucial for colorectal differentiation by regulating the expression of structural proteins important for epithelial differentiation, including villin, cytokeratin 20, and FABP1. Exogenous expression of CDX1 in poorly differentiated cell lines that do not express endogenous CDX1 induces lumen formation in 3D cell cultures. CDX1 is upregulated in Barrett’s metaplasia of the esophagus and transgenic Cdx1 expression in mouse gastric epithelium causes intestinal transdifferentiation. CDX1 is transcriptionally silenced in many colorectal cancers due to promoter methylation.

Specificity

Clone 123a detected the target band in CDX1-expressing colorectal cancer (CRC) cells, but not in non-CDX1-expressing CRC cells. Antibody blocking with the immunogen peptide, but not with a C-terminal peptide, prevented the detection of the target band (Chan, C.W., et al. (2009). Proc. Natl. Acad. Sci .U. S. A. 106(6):1936-1941). Immunogen sequence is not present in spliced isoform 2 reported by UniProt (P47902-2). Clone C is known to exhibit cross-reactivity toward an unidentified protein species in fibroblasts and smooth muscle cells. While such cross-reactivity is often not seen or extremely weak in epithelial cells, immunoreactivity by this clone must be interpreted carefully when handling samples of non-epithelial origins.

Immunogen

Epitope: N-terminal region.

Linear peptide corresponding to a sequence from the N-terminal region of human Cdx1.

Application

Anti-Cdx1 Antibody, clone 123a is an antibody against Cdx1 for use in Western Blotting, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

Western Blotting Analysis: 1.0 µg/mL from a representative lot detected CdX1 in 10 µg of human stomach tissue lysate.
Western Blotting Analysis: A representative lot detected the target band in CDX1-expressing colorectal cancer (CRC) cells (HT55, LS174T, RCM-1, SK-CO-1), but not in non-Cdx1-expressing CRC (DLD-1, HCT116, SW48, RKO) cells (Chan, C.W., et al. (2009). Proc. Natl. Acad. Sci .U. S. A. 106(6):1936-1941).
Western Blotting Analysis: A representative lot detected the endogenously expressed CDX1 in HCT116 human colorectal cancer (CRC) cells. Antibody blocking with the immunogen peptide, but not with a C-terminal peptide, prevented the target band detection (Chan, C.W., et al. (2009). Proc. Natl. Acad. Sci .U. S. A. 106(6):1936-1941).
Immunocytochemistry Analysis: A representative lot detected a downregulated CDX1 immunoreactivity among SW1222 and LS180 colorectal cancer (RC) cell colonies following prolyl-hydrolase inhibitor DMOG (Cat. No. 400091) treatment as a result of enhanced normoxia HIF-α transcription activity (Ashley, N., et al. (2013). Cancer Res. 73(18):5798-5809).
Immunocytochemistry Analysis: A representative lot detected a high expression of the enterocyte differentiation marker CDX1 among colonies formed from colorectal cancer (RC) cells (SW1222, LS180, CCK-81) under normoxia condition, while a much lower Cdx1 immunostaining was seen among the colonies formed under hypoxia condition (Yeung, T.M., et al. (2011). Proc. Natl. Acad. Sci. U. S. A. 108(11):4382-4387).
Immunocytochemistry Analysis: A representative lot immunostained LS174T and CDX1-transfected HCT116 colorectal cancer (CRC) cells. No staining was seen among mock-transfected HCT116 cells or CDX1 shRNA-transfected LS174T cells (Chan, C.W., et al. (2009). Proc. Natl. Acad. Sci. U. S. A. 106(6):1936-1941).
Chromatin Immunoprecipitation ChIP) Analysis: A representative lot detected CDX1 occupancy at the KRT20 promoter site using chromatin preparation from HT55 human colorectal cancer (CRC) cells (Chan, C.W., et al. (2009). Proc. Natl. Acad. Sci. U. S. A. 106(6):1936-1941).

Quality

Evaluated by Western Blotting in human Caco-2 colorectal cancer cell lysate.

Western Blotting Analysis: 1.0 µg/mL of this antibody detected Cdx1 in 10 µg of human Caco-2 colorectal cancer cell lysate.

Target description

~28-32 kDa observed. 28.14 kDa calculated.

Physical form

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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