Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-80611

Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 2D3

Format

Azide and BSA Free
View all Carbonic Anhydrase IX/CA9 Primary Antibodies »

Product Specifications

Immunogen

This Carbonic Anhydrase IX/CA9 Antibody (2D3) was made to a purified recombinant fragment of human Carbonic Anhydrase IX expressed in E. coli [UniProt# Q16790].

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

50 kDa.
Disclaimer 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.

Scientific Data Images for Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free

Western Blot: Carbonic Anhydrase IX/CA9 Antibody (2D3)Azide and BSA Free [NBP2-80611]

Western Blot: Carbonic Anhydrase IX/CA9 Antibody (2D3)Azide and BSA Free [NBP2-80611]

Western Blot: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - Carbonic Anhydrase IX mouse antibody against Hela (1) and A549 (2) cell lysates. Bands were detected at a molecular weight of approximately 50 kDa in both cell lines. Image from the standard format of this antibody.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - Mouse choroid stained with anti-Carbonic Anhydrase IX antibody. Image from verified customer review. Image from the standard format of this antibody.
Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - Paraffin-embedded lung tissues (left) and colonic tissues (right) using Carbonic Anhydrase IX mouse antibody with DAB staining. Image from the standard format of this antibody.
Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - A surface stain was performed on A431 cells with Carbonic Anhydrase IX/CA9 NBP1-51691 (blue) and a matched isotype control NBP2-27287 (orange). Cells were incubated in an antibody dilution of 1 ug/mL for 20 minutes at room temperature, followed by Dylight488-conjugated anti-mouse secondary antibody. Image from the standard format of this antibody.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-CA9 (2D3) [NBP1-51691] at a 1:200 dilution overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.
Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611]

Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free [NBP2-80611] - Analysis of NTERA-2 cells using Carbonic Anhydrase IX mouse mAb (green) and negative control (purple). Image from the standard format of this antibody.

Applications for Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free

Application
Recommended Usage

ELISA

1:10000

Flow (Intracellular)

1 ug/ml

Flow Cytometry

1:200 - 1:400

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Paraffin

1:200 - 1:1000

Western Blot

1:2000
Application Notes
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.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Carbonic Anhydrase IX/CA9

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

Alternate Names

CA9, G250, MN, RCC

Gene Symbol

CA9

Additional Carbonic Anhydrase IX/CA9 Products

Product Documents for Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for Carbonic Anhydrase IX/CA9 Antibody (2D3) - Azide and BSA Free

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.

Related Research Areas

Cancer Biomarkers

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Protocols

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