Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, CyTOF-reported

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry, CyTof, IF/ICC, IF/IHC, Lateral Flow Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Carbonic Anhydrase IX/CA9 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase IX
Pro59-Asp414
Accession # Q16790

Specificity

Detects human Carbonic Anhydrase IX (CA9) in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse CA9 is observed and less than 1% cross-reactivity with recombinant human CA1, 3, 4, 8, 10, 12, and 14 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Carbonic Anhydrase IX/CA9 Antibody

Detection of Human Carbonic Anhydrase IX/CA9 antibody by Western Blot.

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 antibody in A431 Human Cell Line by Immunocytochemistry (ICC).

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 antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P).

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 by Western Blot

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 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

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

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.

Applications for Human Carbonic Anhydrase IX/CA9 Antibody

Application
Recommended Usage

CyTOF-reported

Chevrier, S. et al. (2018) Cell Systems. 6: 612. Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: U87-MG cells

Immunocytochemistry

3-15 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Carbonic Anhydrase IX (Catalog # 2188-CA), see our available Western blot detection antibodies

Western Blot

1 µg/mL
Sample: U‑87 MG human glioblastoma/astrocytoma cell line

Reviewed Applications

Read 2 reviews rated 5 using AF2188 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Carbonic Anhydrase IX/CA9

Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO2 + H2O = HCO3- + H+, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (1-3). Topics in the CA meeting (6th International Conference on the CAs, June 20-25, 2003, in Slovakia) ranged from use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. CA9, also known as membrane antigen MN and renal cell carcinoma (RCC)-associated antigen G250, is a transmembrane enzyme expressed primarily in carcinoma cells. It is one of the best markers for hypoxia and for RCC (4, 5). Recombinant human CA9 corresponds to the extracellular portion of human CA9.

References

  1. Pastorek, J. et al. (1994) Oncogene 9: 2877.
  2. Opavsky, R. et al. (1996) Genomics 33: 480.
  3. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  4. Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
  5. Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.

Alternate Names

CA9, G250, MN, RCC

Entrez Gene IDs

768 (Human); 230099 (Mouse)

Gene Symbol

CA9

UniProt

Q16790

Additional Carbonic Anhydrase IX/CA9 Products

Product Documents for Human Carbonic Anhydrase IX/CA9 Antibody

Certificate of Analysis

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Product Specific Notices for Human Carbonic Anhydrase IX/CA9 Antibody

For research use only

Related Research Areas

Cancer Biomarkers

Citations for Human Carbonic Anhydrase IX/CA9 Antibody

Customer Reviews for Human Carbonic Anhydrase IX/CA9 Antibody (2)

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  • Human Carbonic Anhydrase IX/CA9 Antibody
    Name: Santhosh Sivajothi
    Application: Immunohistochemistry
    Sample Tested: Renal cancer tissue
    Species: Human
    Verified Customer | Posted 05/21/2018
    Immunohistochemistry was performed on FFPE tissue section. Heat induced antigen retrieval was performed by heating section in citrate buffer (pH 6) at 95C for 20 minutes.
    Human Carbonic Anhydrase IX/CA9 Antibody AF2188
  • Human Carbonic Anhydrase IX/CA9 Antibody
    Name: Santhosh Sivajothi
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Renal Cell Carcinoma tissues
    Species: Human
    Verified Customer | Posted 05/18/2018
    Heat mediated antigen retrieval performed in citrate buffer (pH 6) for 20 minutes.
    Human Carbonic Anhydrase IX/CA9 Antibody AF2188

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