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Carbonic Anhydrase IX/CA9 Antibody - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NB100-417

Novus Biologicals, part of Bio-Techne
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NB100-417
NB100-417SS

Key Product Details

Validated by

Knockout/Knockdown, Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Plant

Cited:

Human, Mouse, Rat, Plant

Applications

Validated:

Chromatin Immunoprecipitation (ChIP), Dual RNAscope ISH-IHC, ELISA, Flow Cytometry, Gel Super Shift Assays, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Microarray, Proximity Ligation Assay, Simple Western, Western Blot

Cited:

Block/Neutralize, Chemotaxis, ELISA, EMSA, IF/ICC, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Microarray, Proximity Ligation Assay, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Specifications

Immunogen

This Carbonic Anhydrase IX/CA9 Antibody was made from a synthetic peptide from the C-terminal sequence of human Carbonic Anhydrase IX (within residues 400-459) [UniProt# Q16790].

Localization

Membrane.

Marker

Hypoxia Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

55 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 - BSA Free

Immunohistological Detection of Carbonic Anhydrase IX/CA9 in SCCNij202 Tumors in Mice

Immunohistological Detection of Carbonic Anhydrase IX/CA9 in SCCNij202 Tumors in Mice

CA9 expressed in hypoxic regions as assessed by pimonidazole staining (A), but also observed in non-pimonidazole areas (B). Most of CA9 expressed in pimonidazole positive regions (C). Proliferation (BrdUrd) & apoptosis (caspase-3) in relation to CA9 expression shown in figure D & E. Red, CA9; Green, pimonidazole (A-B), BrdUrd (D) or caspase-3 (E); Yellow, overlap of CA9 (red) & pimonidazole (green); Light blue, vessels. Magnification 100x. Scale bars represent 100 um. Closed circles represent CA9 expression in pimonidazole positive regions; open circles represent CA9 expression in pimonidazole -ve regions. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0108068), licensed under a CC-BY license.
Dual RNAscope ISH-IHC Analysis of Carbonic Anhydrase IX/CA9 in Paraffin Embedded Human Stomach

Dual RNAscope ISH-IHC Analysis of Carbonic Anhydrase IX/CA9 in Paraffin Embedded Human Stomach

Formalin-fixed paraffin-embedded tissue sections of human stomach were probed for Carbonic Anhydrase IX/CA9 mRNA (ACD RNAScope Probe, catalog # 559348; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus catalog # NB100-417) at 1:1000 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to glandular cells.
Western Blot Detection of Carbonic Anhydrase IX/CA9 in Multiple Human Cell Lysates

Western Blot Detection of Carbonic Anhydrase IX/CA9 in Multiple Human Cell Lysates

Analysis on various human cell lysates. Specific bands were detected for Carbonic Anhydrase IX/CA9 in HEK-293 and A498 cell lines at a molecular weight of approximately 50 kDa. WB image submitted by a verified customer review.

Applications for Carbonic Anhydrase IX/CA9 Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

Dual RNAscope ISH-IHC

1:1000

ELISA

reported in scientific literature (PMID 19963243)

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

2 - 5 ug/ml

Immunohistochemistry

1:200 - 1:500

Immunohistochemistry-Frozen

1:200 - 1:500

Immunohistochemistry-Paraffin

1:200 - 1:500

Immunoprecipitation

1:10 - 1:500. Use reported in multiple pieces of scientific literature

Microarray

reported in scientific literature (PMID 31955345)

Simple Western

1:50

Western Blot

1 - 3 ug/ml
Application Notes
In Western blot a band is observed approx. 53 kDa. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 7 reviews rated 4.4 using NB100-417 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at -20C. 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 - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Carbonic Anhydrase IX/CA9 Antibody - 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.

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