Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15))

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) [NBP2-15199] - FFPE human renal cell carcinoma stained with RCC Ab (66.4.C2). Note cytoplasmic & cell surface staining of tumor cells.

• Reactivity: Hu, Eq
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-P, IP
• Clone: 66.4.C2 (PN-15)
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)) Summary

• Immunogen: Fresh, normal human renal cortical tissue homogenate was used to generate the Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15)).
• Localization: Cell surface and cytoplasmic
• Specificity: Recognizes a glycoprotein of ~200kDa, identified as carbonic anhydrase IX (CAIX/gp200). Its epitope resides in the carbohydrate domain of gp200. It shows no significant cross-reactivity with other carbohydrate determinants, such as the Lewis blood group antigens, epithelial membrane antigen, HMFG, and AB blood group antigens. In normal kidney, gp200 is localized along the brush border of the pars convoluta and pars recta segments of the proximal tubule, as well as focally along the luminal surface of Bowman's capsule adjoining the outgoing proximal tubule. Reportedly, gp200 is expressed by 93% of primary and 84% of metastatic renal cell carcinomas. This MAb may be useful in the investigations of carcinomas of proximal nephrogenic differentiation especially those showing tubular differentiation.
• Isotype: IgG2b Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: CA9
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5-1ug/ml
  • ELISA 1-5ug/ml for coating
  • Flow Cytometry 0.5-1ug/million cells
  • Immunocytochemistry/Immunofluorescence 1-2ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 0.5-1ug/ml
  • Immunoprecipitation 1-2ug/500ug protein lysate
• Application Notes: Tripsin (10 min at 37C) is recommend for antigen retrieval (Bayder & Aydin, 2008). The 7mL size is a pre-diluted size and no additional dilutions are required before using this item for the intended application.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein G purified

Alternate Names for Carbonic Anhydrase IX/CA9 Antibody (66.4.C2 (PN-15))

• 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

Background

Carbonic anhydrase alpha, isozyme IX, belongs to a family of zinc-containing metalloproteins which hydrate carbon dioxide to generate bicarbonate ions and protons (1). This main catalytic function allows carbonic anhydrase IX to participate in cellular pH regulation. The large family of carbonic anhydrase metalloproteins includes three major classes which have been identified based on sequence and structure analysis. The alpha class is a monomer found in mammals. The beta class may occur as a dimer, tetramer, hexamer or octamer and is found in plants, algae, and bacteria. Lastly, the gamma class is a trimer found in bacteria and represents the most ancient carbonic anhydrase. These three classes of carbonic anhydrase enzymes lack sequence or structural similarities, but all share a conserved active site zinc atom (1).

Carbonic anhydrase IX (theoretical molecular weight 50kDa) belongs to the monomeric alpha class and is a single pass-transmembrane protein with two extracellular domains which serve catalytic and cell adhesion functions (2, 3). By cooperating with sodium bicarbonate cotransporters (NBC), lactate and proton exporting monocarboxylic acid transporters (MCT), and a sodium/hydrogen exchanger (NHE), carbonic anhydrase IX is involved in pH regulation across the cell membrane. This functional property protects cancer cells from intracellular acidification and partly explains the role of carbonic anhydrase IX in cancer cell survival and proliferation. In contrast, the pH regulating activity of carbonic anhydrase IX induces extracellular acidification, which has been implicated in epithelial to mesenchymal transition (EMT) and promoting cancer invasion. Carbonic anhydrase IX is frequently overexpressed in cancer cells (e.g., colorectal-, breast-, lung-carcinoma and brain tumors), an effect promoted by hypoxia within the tumor microenvironment (4). An exception are tumors carrying pVHL inactivating mutations, such as clear cell renal cell carcinoma (ccRCC), where HIF-alpha is stabilized due to dysfunctional proteasomal targeting and can induce HRE (Hypoxia Response Element) containing genes even under physiological normoxia (5). Carbonic anhydrase IX may be detected by immunostaining in tumors, which is found in association with necrotic tissue and metastatic cells. Because the expression of carbonic anhydrase IX correlates with both tumor grade and stage, analysis of its expression in tumors serves as a prognostic factor (4, 6).

References
1. Tripp, B. C., Smith, K., & Ferry, J. G. (2001). Carbonic Anhydrase: New Insights for an Ancient Enzyme. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.R100045200

2. Nishimori, I., & Onishi, S. (2001). Carbonic anhydrase isozymes in the human pancreas. Digestive and Liver Disease. https://doi.org/10.1016/s1590-8658(01)80138-9

3. Zavadova, Z., & Zavada, J. (2005). Carbonic anhydrase IX (CA IX) mediates tumor cell interactions with microenvironment. Oncology Reports. https://doi.org/10.3892/or.13.5.977

4. Pastorekova, S., & Gillies, R. J. (2019). The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond. Cancer and Metastasis Reviews. https://doi.org/10.1007/s10555-019-09799-0

5. Haase, V. (2009). The VHL Tumor Suppressor: Master Regulator of HIF. Current Pharmaceutical Design. https://doi.org/10.2174/138161209789649394

6. Young, J. R., Coy, H., Kim, H. J., Douek, M., Sisk, A., Pantuck, A. J., & Raman, S. S. (2018). Association of the gross appearance of intratumoral vascularity at MDCT with the carbonic anhydrase IX score in clear cell renal cell carcinoma. American Journal of Roentgenology. https://doi.org/10.2214/AJR.18.19725

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Carbonic Anhydrase IX/CA9 Antibody (NBP2-15199)(5)

Publications using NBP2-15199

• Murakami A, Gomi K, Tanaka M et al. Unilateral glomerulocystic kidney disease associated with tuberous sclerosis complex in a neonate. Pathol Int. 2012 Mar [PMID: 22360510] (IHC-P, Human)
  
  Details:
  IHC (P): Table 1 (kidney mass from a human neonate).
• Chen CJ, Hsu HT, Lin MT et al. Renal cell carcinoma with t(X;17)(p11.2;q25) in a 5-year-old Taiwanese boy: a case report and review of the literature. Pathol Oncol Res. 2012 Jan [PMID: 21221873] (IHC-P, Human)
  
  Details:
  IHC(P): human renal tumor (results described).
• Baydar D, Aydin O. Confusing cases: clear cell but not renal cell lesions in kidney. Pathol Int. 2008 Nov [PMID: 18844937] (IHC-P, Human)
  
  Details:
  IHC (P): Table 1 (human kidney lesions).
• Ingold B, Wild PJ, Nocito A et al. Renal cell carcinoma marker reliably discriminates central nervous system haemangioblastoma from brain metastases of renal cell carcinoma. Histopathology. 2008 May [PMID: 18393979] (IHC-P, Human)
  
  Details:
  IHC(P): human renal cell carcinoma and their metastases, haemangioblastoma (results described).
• Swain JM, Pirie RS, Hudson NP et al. Insulin-like growth factors and recurrent hypoglycemia associated with renal cell carcinoma in a horse. J Vet Intern Med. 2005 Jul-Aug [PMID: 16095185] (IHC-P)
  
  Details:
  IHC (P), Fig 4 (mass of unknown origin in a horse). Note: The antibody helped define the mass as a renal cell carcinoma.

FAQs for Carbonic Anhydrase IX/CA9 Antibody (NBP2-15199). (Showing 1 - 2 of 2 FAQs).

1. I have a couple of questions regarding the Carbonic Anhydrase IX Antibody (NBP2-15199) listed on your website. We are wondering if this antibody is carrier-free, and if not, whether you have a carrier-free version?
   • The buffer for NBP2-15199 contains PBS with 0.05% BSA and 0.05% Sodium Azide. Please see catalog# NBP2-33152 for a carrier free and preservative free version of this antibody.
2. What is the predicted theoretical molecular weight of Carbonic Anhydrase IX/CA9 antibodies?
   • The TMW for Carbonic Anhydrase IX/CA9 antibodies is 50 kDa. Please note the observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Bioinformatics

• Gene Symbol: CA9
• Uniprot:
  • Q16790()