Anti-CDX2 Rabbit Monoclonal Antibody

Western blot analysis of CDX2 using anti-CDX2 antibody (M00877-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human 293T whole cell lysates,
Lane 2: human Caco-2 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CDX2 antigen affinity purified monoclonal antibody (Catalog # M00877-1) at 1:1000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:500 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CDX2 at approximately 37 kDa. The expected band size for CDX2 is at 34 kDa.

All lanes use the Antibody at 1:3K dilution for 1 hour at room temperature.

IHC analysis of CDX2 using anti-CDX2 antibody (M00877-1).
CDX2 was detected in a paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CDX2 Antibody (M00877-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

The distribution of Lgr5 + ISCs in the intestinalmucosa and the subcellular localization and relative expression level detection of epithelial function proteins CDX2 and villin in the intestinal mucosa of IBD at 7 days after termination of DSS administration. (A) The Lgr5 + ISCs (brown) in the small intestinal mucosa: (A1) the normal group, the villi and the crypts were arranged compactly, and Lgr5 + ISCs were observed in the crypts; (A2) the DSS group, the villi and the crypts were scattered, with few Lgr5 + ISCs; (A3) the DSS + B. subtilis -fermented milk group, there were more Lgr5 + ISCs in villi and crypts compared with those in the DSS group. (B) The Lgr5 + ISCs (brown) in the colonic mucosa: (B1) the normal group, the glands were arranged compactly, and there were large amounts of Lgr5 + ISCs at the bottom of the glands; (B2) the DSS group: the ulcers were replaced by scars. No Lgr5 + ISCs were observed in the scars; (B3) the DSS + B. subtilis -fermented milk group: the colonic epithelium was integrated, with some regenerated glands. A number of Lgr5 + ISCs were observed at the bottom of the regenerated glands. (C) The CDX2 was localized in the epithelial cellular nuclei (brown) by immunohistochemistry staining in the small intestinal mucosa: (C1) the normal group: the villi and the crypts were arranged compactly, and CDX2 + epithelial cells were observed on the surface of the villi and the crypts; (C2) the DSS group: the villi and the crypts were scattered, and few CDX2 + epithelial cells were observed on the surface of the crypt and the villi; (C3) the DSS + B. subtilis -fermented milk group: more villi and crypts were observed in comparison with the DSS group, and there were more CDX2 + epithelial cells covering the villi and crypts. (D) The CDX2 was localized in the epithelial cellular nuclei (brown) by immunohistochemistry staining in the colonic mucosa: (D1) the normal group: the colonic glands were arranged compactly, and CDX2 + epithelial cells were observed on the surface of the glands; (D2) the DSS group: the glands were scattered, and few CDX2 + epithelial cells were observed in the scar; (D3) the DSS + B. subtilis -fermented milk group: more colonic glands were observed in comparison with the DSS group, and there were more CDX2 + epithelial cells in the glands. (E) The Mucin2 was localized in the cytoplasm of the goblet cells (brown) by immunohistochemistry staining in the small intestinal mucosa: (E1) the normal group, a number of Mucin2 + goblet cells observed in the epithelium; (E2) the DSS group: only few Mucin2 + goblet cells were observed in the remaining villi and crypts; (E3) the DSS + B. subtilis -fermented milk group: more Mucin2 + goblet cells were observed in the recovered mucosa. (F) The Mucin2 was localized in the cytoplasm of the goblet cells (brown) by immunohistochemistry staining in the colonic mucosa: (F1) the normal group, large amounts of Mucin2 + goblet cells were observed in the mucosa; (F2) the DSS group: only few Mucin2 + goblet cells were observed in the scars; (F3) the DSS + B. subtilis -fermented milk group: more Mucin2 + goblet cells were observed in the recovered colonic mucosa. (G,H) Western blotting was applied for detection of the relative expression level of Lgr5, CDX2, and Mucin2 in the samples containing equivalent ileum and colon. The expression level of Lgr5, CDX2, and Mucin2 in the DSS group was significantly lower than that of the normal (control) group. The expression level of Lgr5, CDX2, and Mucin2 in the DSS + B. subtilis -fermented milk (FM) group was significantly higher than that of the DSS group ( n = 5, ** represents p < 0.01).
Index in PubMed under a CC BY license. PMID: 33519783

IHC analysis of CDX2 using anti-CDX2 antibody (M00877-1).
CDX2 was detected in a paraffin-embedded section of mouse colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CDX2 Antibody (M00877-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Immunohistochemical analysis of paraffin-embedded human colon, using CDX2 Antibody.