Human Carbonic Anhydrase IX/CA9 Antibody

Catalog # Availability Size / Price Qty
AF2188
AF2188-SP
Human Carbonic Anhydrase IX/CA9 Antibody in Western Blot
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Product Details
Citations (4)
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Reviews (1)

Human Carbonic Anhydrase IX/CA9 Antibody Summary

Species Reactivity
Human
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.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase IX
Pro59-Asp414
Accession # Q16790
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
HeLa human cervical epithelial carcinoma cell line
Immunohistochemistry
5-15 µg/mL
See below
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with Recombinant Human Carbonic Anhydrase IX (Catalog # 2188-CA), see our available Western blot detection antibodies
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.
 
Immunocytochemistry
3-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

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

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 (Catalog # 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.

Immunocytochemistry Carbonic  Anhydrase IX/CA9 in A431 Human Cell Line. View Larger

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; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Carbonic Anhydrase IX/CA9 in Human Colon Cancer Tissue. View Larger

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; Catalog # 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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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
  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.
Entrez Gene IDs
768 (Human); 230099 (Mouse)
Alternate Names
CA9; CAIX; CA-IX; Carbonate dehydratase IX; carbonic anhydrase 9; Carbonic Anhydrase IX; carbonic dehydratase; EC 4.2.1.1; G250; Membrane antigen MN; MN; P54/58N; PMW1; RCC; RCC-associated antigen G250; RCC-associated protein G250; Renal cell carcinoma-associated antigen G250

Product Datasheets

Citations for Human Carbonic Anhydrase IX/CA9 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Activation of the hypoxia pathway in breast cancer tissue and patient survival are inversely associated with tumor ascorbate levels
    Authors: EJ Campbell, GU Dachs, HR Morrin, VC Davey, BA Robinson, MCM Vissers
    BMC Cancer, 2019;19(1):307.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  2. Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts
    Authors: EE Hedlund, PC McDonald, O Nemirovsky, S Awrey, LDE Jensen, S Dedhar
    Cancers (Basel), 2019;11(7):.
    Species: Xenograft
    Sample Types: Whole Tissue
    Applications: IHC
  3. Compensation of Signal Spillover in Suspension and Imaging Mass Cytometry.
    Authors: Chevrier S, Crowell H, Zanotelli V, Engler S, Robinson M, Bodenmiller B
    Cell Syst, 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: CyTof
  4. The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion
    Authors: M Swayampaku, PC McDonald, M Vallejo, E Coyaud, SC Chafe, A Westerback, G Venkateswa, J Shankar, G Gao, EMN Laurent, Y Lou, KL Bennewith, CT Supuran, IR Nabi, B Raught, S Dedhar
    Oncogene, 2017;36(45):6244-6261.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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Human Carbonic Anhydrase IX/CA9 Antibody
By Anonymous on 05/21/2018
Application: IHC Sample Tested: Renal cancer tissue Species: Human

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.