Human Carbonic Anhydrase IX/CA9 Antibody

Detection of Human Carbonic Anhydrase IX/CA9 by Western Blot. Western blot shows lysates of U-87 MG human glioblastoma/ astrocytoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Carbonic Anhydrase IX/CA9 at approximately 58 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Carbonic Anhydrase IX/CA9 in A431 Human Cell Line. Carbonic Anhydrase IX/CA9 was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Carbonic Anhydrase IX/CA9 in Human Colon Cancer Tissue. Carbonic Anhydrase IX/CA9 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Carbonic Anhydrase IX/CA9 in U87-MG cells cells by Flow Cytometry. U87-MG cells were stained with Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Detection of Carbonic Anhydrase IX/CA9 by Western Blot Beta-catenin enhances the breast cancer stem cell phenotype in response to hypoxia independently of its nuclear transcriptional activity. A, Western analysis (WB) of beta-catenin, SNAI2 and CA9 protein levels in Ctrl/shBeta MCF7 cells upon Nor/1%pO2 conditions; B, MS forming assay in stable beta-catenin silenced (shBeta) MCF7 cells upon Nor/1%pO2 conditions; C, Cytofluorimetric analysis of CD44high/CD24low stem/progenitor population in ctrl/shBeta MCF7 cells upon Nor/1%pO2 conditions; D, Real-Time PCR analysis of ESR1 mRNA level in Ctrl/shBeta MCF7 cells upon Nor/1%pO2 conditions; E, Immunofluorescence (IF) analysis of Beta-catenin in Nor/1%pO2 MCF7 cells; F, WB analysis of beta-catenin in Nor/1%pO2 MCF7 cells cytoplasmic and nuclear fractions (lamin B and beta-tubulin were used as fractionation controls); G, beta-catenin/TCF transcriptional reporter (TOPFLASH) assay in MCF7 cells and T-MS under Nor/1%pO2. Data are presented as mean +/- s.d.; p values refers to t test. n=3, unless otherwise specified. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24260469), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Carbonic Anhydrase IX/CA9 by Western Blot In normoxic basal-like breast cancer cells, cytoplasmic beta-catenin promotes stem cell features in vitro and tumor growth in vivo via constitutive stabilization of CA9 and SNAI2 mRNAs.A, Beta-catenin IF analysis in luminal MCF7 cells and in basal-like MDA-MB-468 and MDA-MB-231 cells; B, Cytofluorimetric analysis of the CD44high/CD24low population in ctrl/shBeta MDA-MB-468 and MDA-MB-231 cells; C, Ctrl/shBeta MDA-MB-468 10-weeks mammary fat pad xenograft assay (n=5, each group); representative pictures of xenograft tissues hematoxylin-eosin staining are include; D, WB analysis of CA9, SNAI2 and beta-catenin protein levels in ctrl/shBeta normoxic MDA-MB-468 and MDA-MB-231 cells; E, CA9 and SNAI2 mRNA stability assay following inhibition of Polymerase 2 transcriptional activity by actinomycin D (100ng/ml) in Ctrl/shBeta MDA-MB-468 and MDA-MB-231 cells; F, Four weeks growth curve of ctrl/shSNAI2 MDA-MB-231 subcutaneous xenograft assay (n=6, each group); G, Hematoxylin-eosin, ESR1 and CDH1 immunohistochemical stainings in xenograft tissue sections. Data are presented as mean +/- s.d.; p values refers to t test. n=3, unless otherwise specified. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24260469), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Carbonic Anhydrase IX/CA9 by Immunohistochemistry Sunitinib induces hypoxia and Carbonic Anhydrase IX (CAIX) expression in primary Triple Negative Breast Cancer (TNBC) tumors. (a) Images of MDA-MB-231 LM2-4Luc+ primary tumor tissue sections harvested at increasing tumor volumes from animals administered either vehicle or 60 mg/kg sunitinib and immunohistochemically stained for the indicated markers. Scale bars: upper panels, 1 mm; lower panels, 200 μm. (b to e) Image-based quantification of (b) CD31+ blood vessels, (c) CAIX expression, (d) pimonidazole and (e) proliferation. Data show the mean ± standard error of the mean (SEM). n = 3 animals/group with 10 images/animal. * p < 0.05, *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31319613), licensed under a CC-BY license. Not internally tested by R&D Systems.