Carbonic Anhydrase IX/CA9 Antibody Summary
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].
Immunogen affinity purified
Test in a species/application not listed above to receive a full credit towards a future purchase.
- Western Blot 1 - 3 ug/ml
- Simple Western 1:50
- Chromatin Immunoprecipitation 1:10 - 1:500
- Flow Cytometry 1:1000
- Gel Super Shift Assays
- Immunocytochemistry/Immunofluorescence 2 - 5 ug/ml
- Immunohistochemistry 1:200 - 1:500
- Immunohistochemistry-Frozen 1:200 - 1:500
- Immunohistochemistry-Paraffin 1:200 - 1:500
- Dual RNAscope ISH-IHC 1:1000
- Knockout Validated
Gel Super Shift Assays was reported in scientific literature. In Western blot a band is observed approx. 53 kDa. Use in ELISA reported in scientific literature (PMID 19963243). Use in chromatin immunoprecipitation reported in multiple pieces of scientific literature. Knockout validation (PMID: 31585940). Use in Microarray reported in scientific literature (PMID:31955345). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Reactivity to various species reported in scientific literature (Species - PMID: Human - 31311071, Mouse - 31097477, Rat - 25767292, and Plant - 19278636).
Packaging, Storage & Formulations
Store at -20C. Avoid freeze-thaw cycles.
0.02% Sodium Azide
Immunogen affinity purified
Alternate Names for Carbonic Anhydrase IX/CA9 Antibody
- Carbonate dehydratase IX
- carbonic anhydrase 9
- Carbonic Anhydrase IX
- carbonic dehydratase
- EC 184.108.40.206
- Membrane antigen MN
- RCC-associated antigen G250
- RCC-associated protein G250
- Renal cell carcinoma-associated antigen G250
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).
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
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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Product General Protocols
View specific protocols for Carbonic Anhydrase IX/CA9 Antibody (NB100-417):
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
FAQs for Carbonic Anhydrase IX/CA9 Antibody (NB100-417). (Showing 1 - 9 of 9 FAQs).
How many units do we need to purchase in order to stain 150 formalin fixed, paraffin embedded kidney biopsies? It depends on the amount used per slide, so could you please confirm how much antibody do you use per slide in IHC-P? Also, is the ab compatible with ENVISION FLEX+, HIGH pH, FOR AS?
- Unfortunately there is no number that I can provide to you as it is impossible to know for sure how many slides you will be able to stain with a vial. This will depend on your experimental conditions, your samples and how highly expressed the protein is, and what dilution you end up choosing. This antibody is recommended to be used at a 1:1000 dilution for IHC staining as this has worked well for our lab. You should order a single vial and test to determine what amount will be best for your assay rather than order too much or too little without first working with the antibody. As for your second question, that is not something that our lab has tested with so I am unsure if it is compatible or not. It has a fairly normal range pH in the buffer (7-8) so most likely the functions best around that range and not at a high pH.
Could you please let me know the immunogen peptide location for this Ab?
- The immunogen used to create this antibody was made to a cytoplasmic area of the protein.
I'm going to use your CA-9 Antibody for paraffin-Immunohistochemistry. Do I have to do any special pretreatments?
- We recommend antigen retrieval to ensure the epitope is not masked, and we also recommend that you add a permeablizing agent to access the epitope in the nucleus. A protocol specific for this product can be found under Product PDFs, labelled as "IHC Protocol".
We dilute our antibodies in 5% milk TBS-Tween and usually freeze-thaw them multiple times. Is this possible with this antibody when diluted?
- Freezing is not recommended for NB100-417 under any circumstances. Unfortunately, the antibody would no longer be covered by our guarantee if it was stored at a temperature that did not match our storage instructions. Of course, you are free to test your proposed conditions if you would like, but we would not be able to accept responsibility if the antibody was damaged by this.
Is this antibody is carrier-free? If not, do you have a carrier-free version?
- NB100-417 is carrier protein free, but does contain 0.1% sodium azide. The buffer formulation is: Tris-citrate/phosphate, pH 7-8 with 0.1% sodium azide. The following products are carrier protein and azide free (catalog numbers 23300002, H00000768-B01P and H00000768-D01P). If you would like to use NB100-417, we do offer antibody purification kits to remove unwanted buffer ingredients. See this link here for those products.
I have a couple of questions regarding the Carbonic Anhydrase IX Antibody (NB100-417) listed on your website. We are wondering if this antibody is carrier-free, and if not, whether you have a carrier-free version?
- NB100-417 is carrier protein free but does contain 0.025% sodium azide. The buffer formulation is: PBS with 0.025% sodium azide. The following products are carrier protein and azide free (catalog numbers NBP2-34411, NBP2-34408, and NBP2-33152). If you would like to use NB100-417, we do offer antibody purification kits to remove unwanted buffer ingredients.
I am interested in using the CA9 antibody (NB100-417) that you supply. We dilute our antibodies in 5% milk TBS-Tween and usually freeze-thaw them multiple times. Is this possible with this antibody when diluted?
- I have checked with the lab and they have informed me that freezing is not recommended for NB100-417 under any circumstances. Unfortunately, the antibody would no longer be covered by our guarantee if it was stored at a temperature that did not match our storage instructions. Of course, you are free to test your proposed conditions if you would like, but we would not be able to accept responsibility if the antibody was damaged by this.
Could this antibody (CA9) be safely stored at -80C?
- The recommended storage temperature for NB100-417 is -20C. It may be possible to store this antibody at -80C, but this has not been confirmed by our lab. It is most important to ensure that that antibody does not undergo too many freeze-thaw cycles.
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.
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