Human Carbonic Anhydrase IX/CA9 Antibody
R&D Systems, part of Bio-Techne | Catalog # AF2188
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Mouse, Xenograft
Applications
Validated:
CyTOF-reported, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Immunoprecipitation, Western Blot
Cited:
CyTof, Immunocytochemistry, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase IX
Pro59-Asp414
Accession # Q16790
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 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.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
Sample: U87-MG cells
Immunocytochemistry
3-15 µg/mL
Sample: Immersion fixed A431 human epithelial carcinoma cell line
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
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
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
Sample: U‑87 MG human glioblastoma/astrocytoma cell line
Reviewed Applications
Read 2 reviews rated 5 using AF2188 in the following applications:
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.
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
- Pastorek, J. et al. (1994) Oncogene 9: 2877.
- Opavsky, R. et al. (1996) Genomics 33: 480.
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
- Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.
Alternate Names
CA9, G250, MN, RCC
Gene Symbol
CA9
UniProt
Additional Carbonic Anhydrase IX/CA9 Products
Product Documents for Human Carbonic Anhydrase IX/CA9 Antibody
Product Specific Notices for Human Carbonic Anhydrase IX/CA9 Antibody
For research use only
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