Carbonic Anhydrase IX/CA9 Antibody

Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - IHC analysis of a formain fixed paraffin embedded (FFPE) tissue section of human breast cancer using CAIX antibody at 1:1000 dilution. The primary antibody bound to CAIX antigens in the tissue section was detected using a HRP labeled secondary antibody and DAB reagent. Nuclei of the cells were counterstained with hematoxylin. This CAIX antibody generated an expected cytoplasmic staining of CAIX protein with an intense signal around the cellular membranes in tumor cores. The latter are more likely to be hypoxic in growing tumors which signifies that the observed CAIX staining is specific.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis in 1) HeLa, 2) MDA-MB-231 and 3) A549 whole cell lysates.

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Immunofluorescence of human RCC tumor cryosections using NB100-417 (Panel A). Panel B shows staining with normal rabbit serum.

Immunohistochemistry-Frozen: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of human colon carcinoma, xenografted in mice. Image courtesy of an anonymous product review.

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of HRP conjugate of NB100-417. HEK 293 cells using NB100-417. Panel A shows CAIX-transfected cells (epitope-unmasked with SDS) and panel B shows mock transfected cells.

Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the Biotin conjugate. Staining of Renal carcinoma tissue.

Simple Western: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Simple Western lane view shows a specific band for CAIX in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the HRP conjugate. Analysis in transfected HEK cell lysate. Rabbit IgG was used as a negative control.

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-CAIX at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - An intracellular stain was performed on U-87 MG Cells with NB100-417 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG APC-conjugated Secondary Antibody, (R&D Systems, F0111).

Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Anaysis in transfected HEK cell lysate using NB 100-417. Rabbit IgG was used as a negative control.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis on rat renal cortex.

Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis on various human cell lysates. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the DyLight 488 conjugate of NB100-417. Staining of Carbonic Anhydrase IX (red) in human glioma U87 cells. DAPI counterstains nuclei (blue). Image provided via product review by verified customer.

Immunoblotting: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Immunoblotting was used to determine CAIX/Carbonic Anhydrase IX/CA9 and other antibodies protein levels in total extracts. Cells were transfected using the specified siRNA for 6 days or treated with desferrioxamine (DFO) for 16 hours. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

Immunoblotting: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Iron depletion and nitric oxide stress and Iron depletion induce the truncated VDAC1 form accumulation. (D) Total protein extracts were analyzed by immunoblotting using VDACs poly antibody and anti-HIF-1 alpha and -CAIX antibodies. (E) Immunoblotting was used to analyze total protein extracts using antibodies against the three VDAC isoforms. (F) HeLa cells were grown in hypoxia (Hx, 1% O2) conditions and transfected with iscu- or NC-siRNA for 3 days, or grown in normoxia (Nx, 21% O2), some treated with DFO for 16 h. Western Blots analyzed total proteins using VDACs poly antibody and anti-HIF-1 alpha, -CAIX, -ISCU antibodies. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

Immunoblotting: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Hypoxias impact on proteins of the mitochondrial ISC assembly machinery. (A) Immunoblotting analyzed the total protein extracts from HeLa cells grown in normoxia (Nx, 21% O2) or hypoxia (Hx, 1% O2) conditions using VDACs poly antibody and anti-CAIX, -ISCU, -FXN, -NFS1, -HSC20 antibodies. Beta-Actin was used as loading control. Citation: Ferecatu I, Canal F, Fabbri L, Mazure NM, Bouton C, Golinelli-Cohen M-P (2018) Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1 alpha activation. PLoS ONE 13(3): e0194782.

• Reactivity: Hu, Mu, Rt, Ca, Pl
• Applications: WB, Simple Western, ChIP, ELISA, Flow, GS, IB, ICC/IF, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Carbonic Anhydrase IX/CA9 Antibody Summary

• Immunogen: A synthetic peptide made to a C-terminal sequence of human Carbonic Anhydrase IX (within residues 400-459) [UniProt Q16790]
• Localization: Membrane.
• Marker: Hypoxia Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CA9
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:2000
  • Simple Western 1:50
  • Chromatin Immunoprecipitation 1:10-1:500
  • ELISA
  • Flow Cytometry 1:1000
  • Gel Super Shift Assays
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:1000
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Frozen 1:1000
  • Immunohistochemistry-Paraffin 1:1000
• Application Notes: This Carbonic Anhydrase IX antibody is useful for Western blot, Immunofluorescence/Immunocytochemistry, and Immunohistochemistry. 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.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  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.
• 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.
• Positive Control:

  HeLa Lysate (NB800-PC1)
  Carbonic Anhydrase IX/CA9 Lysate (NBL1-08619)

• Control Peptide:

  Carbonic Anhydrase IX/CA9 Peptide (NB100-417PEP)

Reactivity Notes

Human, dog, rat and mouse (PMID 22842475). Plant reactivity reported in scientific literature (PMID: 19278636)

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.025% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Carbonic Anhydrase IX/CA9 Antibody

• CA9
• CA-IX
• CAIXcarbonic anhydrase 9
• Carbonate dehydratase IX
• Carbonic Anhydrase IX
• carbonic anhydrase IXpMW1
• carbonic dehydratase
• EC 4.2.1.1
• G250
• Membrane antigen MN
• MN
• MNRCC-associated antigen G250
• P54/58N
• RCC
• RCC-associated protein G250
• Renal cell carcinoma-associated antigen G250

Background

Carbonic anhydrase IX (CAIX) is a member of the Carbonic anhydrase family which are enzymes that assists rapid interconversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. They are abundant in all mammalian tissues and because of their functionality, they have become an important diagnostic marker for various cancers, most notably renal cell carcinoma (RCC). Carbonic Anhydrase IX (CA IX) is one of the most hypoxically inducible genes because of its stability and the location of the expressed protein within the membrane. Carbonic anhydrases have a widespread role in regulating pH in normal tissues, by regulating hydrogen ion (H+) flux. The pH is important in cell death under hypoxia, thus a blockade of CA IX (CA9) results in increased cell death under hypoxia. Therefore, CA IX has become a reliable histochemical marker of hypoxia.

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 (NB100-417)(152)

Publications using NB100-417

• Ames S, Pastorekova S, Becker HM. The proteoglycan-like domain of carbonic anhydrase IX mediates non-catalytic facilitation of lactate transport in cancer cells. Oncotarget. Jun 15 2018 [PMID: 29963253] (WB, Human)
• Chen L, Chen L, Huang J et al. Tyrosine kinase inhibitor sunitinib targets the vasculature of clear cell renal cell carcinoma: a morphometrical study of treatment effect. Int J Clin Exp Pathol. 2016 (IHC-P, Human)
• Lambrechts D, Wauters E, Boeckx B et al. Phenotype molding of stromal cells in the lung tumor microenvironment. Nat. Med. Jul 9 2018 [PMID: 29988129] (IHC-P, Human)
• Iwamoto H, Abe M, Yang Y et al. Cancer Lipid Metabolism Confers Antiangiogenic Drug Resistance Cell Metab. May 25 2018 [PMID: 29861385] (ICC/IF)
• Rosenberg T, Aaberg-Jessen C, Petterson SA, Kristensen BW. Heterogenic expression of stem cell markers in patient-derived glioblastoma spheroid cultures exposed to long-term hypoxia CNS Oncol Apr 1 2018 [PMID: 29708435] (IHC, Human)
• Ferecatu I, Canal F, Fabbri L et al. Dysfunction in the mitochondrial Fe-S assembly machinery leads to formation of the chemoresistant truncated VDAC1 isoform without HIF-1a activation. PLoS ONE Mar 29 2018 [PMID: 29596470] (WB, Human)
  
  Details:
  CAIX antibody from Novus was used to look at the effect of desferrioxamine treatment on human cell lines with regards to CAIX protein levels in western blot
• Corbet C, Bastien E, Draoui N et al. Interruption of lactate uptake by inhibiting mitochondrial pyruvate transport unravels direct antitumor and radiosensitizing effects. Nat Commun. Mar 23 2018 [PMID: 29572438] (IHC, Human)
• Young JR, Coy H, Kim HJ et al. Utility of multiphasic multidetector computed tomography in discriminating between clear cell renal cell carcinomas with high and low carbonic anhydrase-IX expression. Abdom Radiol (NY). Mar 8 2018 [PMID: 29520426] (IHC)
• Dreyer-Andersen N, Almeida AS, Jensen P et al. Intermittent, low dose carbon monoxide exposure enhances survival and dopaminergic differentiation of human neural stem cells PLoS ONE 2018 Jan 16 [PMID: 29338033] (ICC/IF, Human)
• Boyd NH, Walker K, Fried J et al. Addition of carbonic anhydrase 9 inhibitor SLC-0111 to temozolomide treatment delays glioblastoma growth in vivo. JCI Insight 2017 Dec 21 [PMID: 29263302] (WB, Human)
• He L, Zhao C, Li Y et al. Antiangiogenic effects of recombinant human endostatin in lung cancers. Mol Med Rep. 2017 [PMID: 29115591] (Mouse)
• Nakamura M, Zhang Y, Yang Y et al. Off-tumor targets compromise antiangiogenic drug sensitivity by inducing kidney erythropoietin production. Proc. Natl. Acad. Sci. U.S.A. 2017 Oct 23 [PMID: 29078273] (Mouse)
• Wang D, Berglund AE, Kenchappa RS et al. BIRC3 is a biomarker of mesenchymal habitat of glioblastoma, and a mediator of survival adaptation in hypoxia-driven glioblastoma habitats Sci Rep 2017 Aug 24 [PMID: 28839258] (WB, Mouse)
• Bucau M, Laurent-Bellue A, Pote N et al. 18F-FDG uptake in well-differentiated neuroendocrine tumors correlates with both Ki-67 and VHL pathway inactivation Neuroendocrinology 2017 Aug 11 [PMID: 28803235] (Human)
• Klameth L, Rath B, Hochmaier M et al. Small cell lung cancer: model of circulating tumor cell tumorospheres in chemoresistance Sci Rep 2017 Jul 13 [PMID: 28706293] (IHC-P, Human)
• Zhang Y, Sun M, Huang G et al. Maintenance of antiangiogenic and antitumor effects by orally active low-dose capecitabine for long-term cancer therapy. Proc. Natl. Acad. Sci. U.S.A. 2017 Jun 27 [PMID: 28607065]
• Takenaka M, Suzuki N, Mori M et al. Iron regulatory protein 2 in ovarian endometrial cysts. Biochem. Biophys. Res. Commun. Apr 24 2017 [PMID: 28450115]
• Bousquet G, El Bouchtaoui M, Sophie T et al. Targeting autophagic cancer stem-cells to reverse chemoresistance in human triple negative breast cancer. Oncotarget. 2017 May 23 [PMID: 28445132] (ICC/IF, Human)
• Himmels P, Paredes I, Adler H et al. Motor neurons control blood vessel patterning in the developing spinal cord. Nat Commun. Mar 6 2017 [PMID: 28262664] (IHC-FR, Mouse)
• Fraga A, Ribeiro R, Coelho A et al. Genetic polymorphisms in key hypoxia-regulated downstream molecules and phenotypic correlation in prostate cancer. BMC Urol 2017 Jan 31 [PMID: 28143503] (Human)
• Bulusu V, Tumanov S, Michalopoulou E et al. Acetate Recapturing by Nuclear Acetyl-CoA Synthetase 2 Prevents Loss of Histone Acetylation during Oxygen and Serum Limitation Cell Rep Jan 17 2017 [PMID: 28099844] (IHC, Human)
• Tsuji T, Kurokawa Y, Chiche J et al. Dissecting the process of activation of cancer-promoting zinc-requiring ectoenzymes by zinc metalation mediated by ZnT transporters J. Biol. Chem Dec 27 2016 [PMID: 28028180] (WB)
• Oh ET, Kim JW, Kim JM et al. NQO1 inhibits proteasome-mediated degradation of HIF-1a. Nat Commun. Dec 14 2016 [PMID: 27966538] (IHC, Mouse)
• Frost J, Galdeano C, Soares P et al. Potent and selective chemical probe of hypoxic signalling downstream of HIF-alpha hydroxylation via VHL inhibition. Nat Commun. Nov 4 2016 [PMID: 27811928] (WB)
• Cauchy F, Mebarki M, Leporq B et al. Strong antineoplastic effects of metformin in preclinical models of liver carcinogenesis. Clin. Sci. Nov 1 2016 [PMID: 27803295]
• Eikrem OS, Strauss P, Beisland C et al. Development and confirmation of potential gene classifiers of human clear cell renal cell carcinoma using next-generation RNA sequencing. Scand J Urol Dec 1 2016 [PMID: 27739342] (IHC, Human)
• Huang Y, Lichtenberger LM, Taylor M et al. Anti-tumor and Anti-angiogenic Effects of Aspirin-PC in Ovarian Cancer Mol. Cancer Ther. Sep 16 2016 [PMID: 27638860] (IHC, Human)
• Kim En, Kim Gh, Kim J et al. A Pyloric Gland-Phenotype Ovarian Mucinous Tumor Resembling Lobular Endocervical Glandular Hyperplasia in a Patient with Peutz-Jeghers Syndrome. J Pathol Transl Med Aug 22 2016 [PMID: 27550049] (IHC-P)
• van Kuijk SJ, Gieling RG, Niemans R et al. The Sulfamate Small Molecule CAIX Inhibitor S4 Modulates Doxorubicin Efficacy. PLoS ONE. Aug 12 2016 [PMID: 27513947] (IHC, Human)
• Hirakawa T, Yashiro M, Doi Y et al. Pancreatic Fibroblasts Stimulate the Motility of Pancreatic Cancer Cells through IGF1/IGF1R Signaling under Hypoxia. PLoS ONE. Aug 4 2016 [PMID: 27487118] (IHC, Human)
• Yoon C, Chang KK, Lee JH et al. Multimodal targeting of tumor vasculature and cancer stem-like cells in sarcomas with VEGF-A inhibition, HIF-1a inhibition, and hypoxia-activated chemotherapy Oncotarget Jul 12 2016 [PMID: 27374091] (Mouse)
• Leek R, Grimes DR, Harris AL, McIntyre A. Methods: Using Three-Dimensional Culture (Spheroids) as an In Vitro Model of Tumour Hypoxia. Adv. Exp. Med. Biol. Jun 21 2016 [PMID: 27325267] (IHC, Human, Mouse)
• He H, Yang Y, Xiang Z et al. A Sensitive IHC Method for Monitoring Autophagy-Specific Markers in Human Tumor Xenografts. J Biomark. Jun 1 2016 [PMID: 27247826] (IHC-P, Human)
• Hecht JL, Zsengeller ZK, Spiel M, Karumanchi SA. Revisiting decidual vasculopathy. Placenta. 2016 Apr 08 [PMID: 27238712] (IHC-P, Human)
• Xu W, Ghosh S, Comhair SA et al. Increased mitochondrial arginine metabolism supports bioenergetics in asthma. J. Clin. Invest. May 23 2016 [PMID: 27214549] (IHC-P, Mouse)
• Badar T, Handisides DR, Benito JM et al. A Phase I Study of Evofosfamide, an Investigational Hypoxia-Activated Prodrug, in Patients with Advanced Leukemia. Am. J. Hematol. May 11 2016 [PMID: 27169385] (IHC-P, Human)
• Faes S, Planche A, Uldry E et al. Targeting carbonic anhydrase IX improves the anti-cancer efficacy of mTOR inhibitors. Oncotarget. Jun 14 2016 [PMID: 27153561] (IHC, Mouse)
• Ono M, Yamada K, Bensaddek D et al. Enhanced snoMEN Vectors Facilitate Establishment of GFP-HIF-1alpha Protein Replacement Human Cell Lines. PLoS ONE. Apr 30 2016 [PMID: 27128805] (WB, Human)
• Eikrem O, Beisland C, Hjelle K et al. Transcriptome Sequencing (RNAseq) Enables Utilization of Formalin-Fixed, Paraffin-Embedded Biopsies with Clear Cell Renal Cell Carcinoma for Exploration of Disease Biology and Biomarker Development PLoS ONE Feb 23 2016 [PMID: 26901863] (IHC-P, Human)
• Radoul M, Chaumeil MM, Eriksson P et al. MR Studies of Glioblastoma Models Treated with Dual PI3K/mTOR Inhibitor and Temozolomide:Metabolic Changes Are Associated with Enhanced Survival. Mol. Cancer Ther. May 1 2016 [PMID: 26883274] (IHC, Mouse)
• Scholz A, Harter PN, Cremer S et al. Endothelial cell-derived angiopoietin-2 is a therapeutic target in treatment-naive and bevacizumab-resistant glioblastoma. EMBO Mol Med. 2016 Jan 05 [PMID: 26666269] (IHC-P, Human)
• Mitsuhashi A, Goto H, Saijo A et al. Fibrocyte-like cells mediate acquired resistance to anti-angiogenic therapy with bevacizumab. Nat Commun. 2015 Dec 04 [PMID: 26635184] (IHC-Fr, Mouse)
• Andre H, Tunik S, Aronsson M, Kvanta A. Hypoxia-Inducible Factor-1alpha Is Associated With Sprouting Angiogenesis in the Murine Laser-Induced Choroidal Neovascularization Model. ARVO Journals. 2015 Oct [PMID: 26501235] (ICC/IF, Mouse)
• Klier M, Jamali S, Ames S et al. Catalytic activity of human carbonic anhydrase isoform IX is displayed both extra- and intracellularly. FEBS J 2016 Jan [PMID: 26470855]
• Islam SS, Mokhtari RB, Akbari P et al. Simultaneous Targeting of Bladder Tumor Growth, Survival, and Epithelial-to-Mesenchymal Transition with a Novel Therapeutic Combination of Acetazolamide (AZ) and Sulforaphane (SFN). Target Oncol 2015 Oct 10 [PMID: 26453055]
• Kinoshita H, Yashiro M, Fukuoka T et al. Diffuse-type Gastric Cancer Cells Switch Their Driver Pathways from FGFR2 Signaling to SDF1/CXCR4 Axis in Hypoxic Tumor Microenvironments. Carcinogenesis 2015 Sep 18 [PMID: 26385890] (IHC-P, Mouse)
• Jamali S, Klier M, Ames S et al. Hypoxia-induced carbonic anhydrase IX facilitates lactate flux in human breast cancer cells by non-catalytic function. Sci Rep 2015 Sep 04 [PMID: 26337752] (WB, ICC/IF, Human)
• Baine MK, Turcu G, Zito CR et al. Characterization of tumor infiltrating lymphocytes in paired primary and metastatic renal cell carcinoma specimens. Oncotarget 2015 Sep 22 [PMID: 26317902]
• Song CW, Lee YJ, Griffin RJ et al. Indirect Tumor Cell Death After High-Dose Hypofractionated Irradiation: Implications for Stereotactic Body Radiation Therapy and Stereotactic Radiation Surgery. Int J Radiat Oncol Biol Phys 2015 Sep 1 [PMID: 26279032]
• Ward C, Meehan J, Mullen P et al. Evaluation of carbonic anhydrase IX as a therapeutic target for inhibition of breast cancer invasion and metastasis using a series of in vitro breast cancer models Oncotarget 2015 Jul 04 [PMID: 26259239]
• Kasashima H, Yashiro M, Kinoshita H et al. Lysyl oxidase is associated with the epithelial-mesenchymal transition of gastric cancer cells in hypoxia. Gastric Cancer 2016 Apr [PMID: 26100130]
• Yoon C, Lee HJ, Park DJ et al. Hypoxia-activated chemotherapeutic TH-302 enhances the effects of VEGF-A inhibition and radiation on sarcomas. Br. J. Cancer. 2015 May 26 [PMID: 26010414] (IHC-P, ICC/IF, Human)
• Dulloo I, Phang BH, Othman R et al. Hypoxia-inducible TAp73 supports tumorigenesis by regulating the angiogenic transcriptome. Nat. Cell Biol. 2015 Apr 01 [PMID: 25774835] (IHC-P, Human)
• Cotroneo E, Ashek A, Wang L et al. Iron Homeostasis and Pulmonary Hypertension: Iron Deficiency Leads to Pulmonary Vascular Remodeling in the Rat. Circ. Res. 2015 Mar 12 [PMID: 25767292] (IHC, Rat)
• Kang HJ, Kim IH, Sung CO et al. Expression of carbonic anhydrase 9 is a novel prognostic marker in resectable hepatocellular carcinoma Virchows Arch. 2015 Jan 11 [PMID: 25577552] (IHC-P, Human)
  
  Details:
  Carbonic Anhydrase IX/CA9 antibody used at 1:1000 dilution for IHC-P staining on formalin-fixed, paraffin-embedded tissue sections/TMAs created from surgically resected hepatocellular carcinoma/HCC and adjacent non-neoplastic liver tissue - 32 minutes of heat-induced epitope retrieval with Cell Conditioning 1 /CC1 buffer, 16 minutes of primary antibody incubation on Ventana's autoimmunostainer/Bench Mark XT automatic immunostaining device, staining development with Ventana's OptiView DAB IHC Detection Kit. Fig 2a-f documents the staining images showing weak, moderate and strong membranous expression levels of carbonic anhydrase 9 protein and the overall data in this study establishes CA9 protein as a poor prognostic factor in resectable HCC patients.
• Mudie S, Bandarra D, Batie M et al. PITX1, a specificity determinant in the HIF-1alpha-mediated transcriptional response to hypoxia. Cell Cycle 2014 [PMID: 25558831] (WB)
• Lee H, Yoon C, Park DJ et al. Inhibition of Vascular Endothelial Growth Factor A and Hypoxia-Inducible Factor 1alpha Maximizes the Effects of Radiation in Sarcoma Mouse Models Through Destruction of Tumor Vasculature. Int. J. Radiat. Oncol. Biol. Phys. 2014 Dec 24 [PMID: 25544668]
• Janmohamed SR, Brinkhuizen T, den Hollander JC et al. Support for the hypoxia theory in the pathogenesis of infantile haemangioma. Clin. Exp. Dermatol. 2014 Dec 16 [PMID: 25511669] (IHC-P, Human)
• Lin XY, Zhang H, Tang N et al. Expression and diagnostic implications of carbonic anhydrase IX in several tumours with predominantly clear cell morphology. Histopathology. 2014 Nov 27 [PMID: 25431204] (IHC-P, Human)
  
  Details:
  Carbonic Anhydrase IX/CA9 antibody used for IHC-P on sections of clear cell renal cell carcinoma (ccRCC; primary and metastatic) and several other type of tumors with predominantly clear cell morphology - tissues previously fixed in 10% neutral buffered formalin and embedded in paraffin, 4um sections, routine IHC-P processing with streptavidin-peroxidase detection system. ccRCC, and breast / pulmonary/hepatic clear cell carcinoma, urothelial carcinoma with clear cell change, clear cell meningioma/ependymoma, haemangioblastoma, and clear cell hidradenoma showed expression of CAIX. See full text for staining images and the expression levels.
• Gellert LL, Mehra R, Chen YB et al. The diagnostic accuracy of percutaneous renal needle core biopsy and its potential impact on the clinical management of renal cortical neoplasms. Arch. Pathol. Lab. Med. 2014 Dec 01 [PMID: 25427045] (IHC-P, Human)
  
  Details:
  Carbonic Anhydrase IX/CA9 antibody used for IHC-P on sections of human renal biopsies having sarcomatoid area with focal necrosis and clear cell renal cell carcinoma (Figure 3.).
• Santos Cd, Tijeras-Raballand A, Serova M et al. Effects of preset sequential administrations of sunitinib and everolimus on tumour differentiation in Caki-1 renal cell carcinoma. Br. J. Cancer. 2014 Nov 25 [PMID: 25422908] (IHC-P, Human)
• Hwa JS, Kwon OJ, Park JJ et al. The prognostic value of immunohistochemical markers for oral tongue squamous cell carcinoma. Eur Arch Otorhinolaryngol. 2014 Aug 29 [PMID: 25169079]
• Chen LQ, Howison CM, Spier C et al. Assessment of carbonic anhydrase IX expression and extracellular pH in B-cell lymphoma cell line models. Leuk. Lymphoma. 2014 Aug 17 [PMID: 25130478]
• Motzer RJ, Hutson TE, Hudes GR et al. Investigation of novel circulating proteins, germ line single-nucleotide polymorphisms, and molecular tumor markers as potential efficacy biomarkers of first-line sunitinib therapy for advanced renal cell carcinoma. Cancer Chemother. Pharmacol. 2014 Aug 07 [PMID: 25100134] (IHC-P, Human)
  
  Details:
  Rabbit polyclonal Carbonic Anhydrase IX/CA-IX antibody used for IHC on formalin-fixed paraffin-embedded (FFPE) blocks or unstained slides from tumor specimens.
• Sadlecki P, Bodnar M, Grabiec M. The Role of Hypoxia-Inducible Factor-1a, Glucose Transporter-1,(GLUT-1) and Carbon Anhydrase IX in Endometrial Cancer Patients. BioMed Research International 3/12/2014 [PMID: 24745019] (IHC-P, Human)
• Heijmen L, Voert EGW, Punt CJA. Monitoring hypoxia and vasculature during bevacizumab treatment in a murine colorectal cancer model. Contrast Media Mol Imaging 2014 May-Jun [PMID: 24700751] (IHC-Fr, Human)
  
  Details:
  Using the Biotin conjugated version of NB100-417, catalog number NB100-417B.
• Kwon OJ, Park JJ, Ko GH et al. HIF-1a and CA-IX as predictors of locoregional control for determining the optimal treatment modality for early-stage laryngeal carcinoma. Head Neck 2/12/2014 [PMID: 24677746] (IHC-P, Human)
• Chen X, Iliopoulos D, Zhang Q et al. XBP1 promotes triple-negative breast cancer by controlling the HIF1a pathway. Nature 4/3/2014 [PMID: 24670641] (IHC-P, Human)
• Morifuji Yoshihiro, Onishi Hideya, Iwasaki Hironori et al. Reoxygenation from chronic hypoxia promotes metastatic processes in pancreatic cancer through the Hedgehog signaling. Cancer Sci. 2014 Feb 11 [PMID: 24397700]
• Polanski Radoslaw, Hodgkinson Cassandra L, Fusi Alberto et al. Activity of the monocarboxylate transporter 1 inhibitor AZD3965 in small cell lung cancer. Clin Cancer Res. 2014 Feb 15 [PMID: 24277449] (IHC)
• Pinato DJ, Tan TM, Toussi ST et al. An expression signature of the angiogenic response in gastrointestinal neuroendocrine tumours: correlation with tumour phenotype and survival outcomes. Br J Cancer. 2013 Nov 14 [PMID: 24231952] (IHC-P, Human)
• Chaudary N, Pintilie M, Schwock J et al. Characterization of the Tumor-Microenvironment in Patient-Derived Cervix Xenografts (OCICx). Cancers (Basel) 2012 [PMID: 24213469] (IHC)
• Zhang M, Boughton P, Rose B et al. The Use of Porous Scaffold as a Tumor Model. Int J Biomater. 2013 [PMID: 24101930] (IHC, Human)
• Wen YA, Stevens PD, Gasser ML et al. Downregulation of PHLPP Expression Contributes to Hypoxia-Induced Resistance to Chemotherapy in Colon Cancer Cells. Mol Cell Biol. 2013 Nov [PMID: 24061475] (WB, Human)
• El-Heliebi A, Kroneis T, Zohrer E et al. Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?. J Transl Med. 2013 Sep 17 [PMID: 24044779] (ICC/IF, Human)
• Fahling M, Mathia S, Paliege A et al. Tubular von Hippel-Lindau Knockout Protects against Rhabdomyolysis-Induced AKI. J Am Soc Nephrol. 2013 Aug 22 [PMID: 23970125]
• Cioffi S, Martucciello S, Fulcoli FG et al. Tbx1 regulates brain vascularization. Hum Mol Genet. 2013 Sep 7 [PMID: 23945394] (IHC-Fr, ICC/IF, Mouse)
• Kumar S, Mokhtari RB, Oliveira ID et al. Tumor Dynamics in Response to Antiangiogenic Therapy with Oral Metronomic Topotecan and Pazopanib in Neuroblastoma Xenografts. Transl Oncol 2013 Aug 1 [PMID: 23908692] (IHC, Human)
• Dubois L, Peeters SG, van Kuijk SJ et al. Targeting carbonic anhydrase IX by nitroimidazole based sulfamides enhances the therapeutic effect of tumor irradiation: A new concept of dual targeting drugs. Radiother Oncol 2013 Jul 9 [PMID: 23849171] (IHC-Fr, Mouse)
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  Details:
  Using the Biotin conjugated version of NB100-417, catalog number NB100-417B.
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• Nelson MP, Christmann BS, Werner JL et al. IL-33 and M2a alveolar macrophages promote lung defense against the atypical fungal pathogen Pneumocystis murina. J Immunol 2011 Feb 15 [PMID: 21220696] (ChIP)
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• Newton IP, Kenneth NS, Appleton PL et al. Adenomatous polyposis coli and hypoxia-inducible factor-1{alpha} have an antagonistic connection. Mol Biol Cell 2010 Nov 1 [PMID: 20844082] (ChIP)
• Pistollato F, Rampazzo E, Persano L et al. Interaction of hypoxia-inducible factor-1a and Notch signaling regulates medulloblastoma precursor proliferation and fate. Stem Cells 2010 Nov [PMID: 20827750] (IHC-P, Human)
• Kato Y, Yashiro M, Noda S et al. Expression of a hypoxia-associated protein, carbonic anhydrase-9, correlates with malignant phenotypes of gastric carcinoma. Digestion 2010 [PMID: 20588040] (IHC, Human)
• O'Toole D, Couvelard A, Rebours V et al. Molecular markers associated with response to chemotherapy in gastro-entero-pancreatic neuroendocrine tumors. Endocr Relat Cancer 2010 Dec [PMID: 20570957] (IHC, Human)
• Stillebroer AB, Mulders PF, Boerman OC et al. Carbonic anhydrase IX in renal cell carcinoma: implications for prognosis, diagnosis, and therapy. Eur Urol 2010 Jul [PMID: 20359812]
• Pistollato F, Abbadi S, Rampazzo E et al. Intratumoral hypoxic gradient drives stem cells distribution and MGMT expression in glioblastoma. Stem Cells 2010 May [PMID: 20309962] (IHC-P, Human)
• Park I, Larson PE, Zierhut ML et al. Hyperpolarized 13C magnetic resonance metabolic imaging: application to brain tumors. Neuro Oncol 2010 Feb [PMID: 20150380] (IHC)
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• Zhou GX, Ireland J, Rayman P et al. Quantification of carbonic anhydrase IX expression in serum and tissue of renal cell carcinoma patients using enzyme-linked immunosorbent assay: prognostic and diagnostic potentials. Urology 2010 Feb [PMID: 19963243] (ELISA, Human)
• Narita T, Yin S, Gelin CF et al. Identification of a Novel Small Molecule HIF-1{alpha} Translation Inhibitor. Clin Cancer Res 9;15(19):6128-6136. 200 [PMID: 19789328] (WB, Human)
• Hong YS, Cho HJ, Kim SY et al. Carbonic anhydrase 9 is a predictive marker of survival benefit from lower dose of bevacizumab in patients with previously treated metastatic colorectal cancer. BMC Cancer 2009 [PMID: 19619339] (IHC, Human)
• Lucio-Eterovic AK, Piao Y, de Groot JF. Mediators of Glioblastoma Resistance Invasion during Antivascular Endothelial Growth Factor Therapy. Clin Cancer Res:1078-0432.CCR-1009-0575. 2009 [PMID: 19567589] (IHC-P, Human)
• Loinard C, Ginouves A, Vilar J et al. Inhibition of Prolyl Hydroxylase Domain Proteins Promotes Therapeutic Revascularization. Circulation;120(1):50-59. 2009 [PMID: 19546390] (WB, Human, Mouse)
• Li Y, Wang H, Oosterwijk E et al. Antibody-specific detection of CAIX in breast and prostate cancers. Biochem Biophys Res Commun 2009 Aug 28 [PMID: 19538935] (IHC-P, Human)
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• Nakao M, Ishii G, Nagai K et al. Prognostic significance of carbonic anhydrase IX expression by cancer-associated fibroblasts in lung adenocarcinoma. Cancer 2009 Jun 15 [PMID: 19365853] (WB, Human)
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Reviews for Carbonic Anhydrase IX/CA9 Antibody (NB100-417) (4)

Average Rating: 4.5

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Mouse.

reviewed by: Anonymous IHC-P Mouse 10/25/2017

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: IHC on parrafin sections (mouse)
• Species: Mouse
• Lot: AD-1

reviewed by: rajesha rupaimoole WB Human 12/10/2014

Summary

• Application: Western Blot
• Sample Tested: A2780 ovarian cancer cells
• Species: Human

Blocking

• Blocking Details: 5% BSA

Details

• Detection Notes: ECL

reviewed by: Xiuquan Luo WB Human 07/21/2014

Summary

• Application: Western Blot
• Sample Tested: 20 ug whole cell lysate
• Species: Human

Blocking

• Blocking Details: 1xPBST with 5% non-fat milk, RT 1 hour

Primary Anitbody

• Dilution Ratio: 1xpBST with 5% non-fat milk, overnight 4C, 1:1000 dilution

Secondary Antibody

• Secondary Description: Goat anti-rabbit IgG-HRP
• Secondary Manufacturer Cat#: sc-2004
• Secondary Concentration: 1:2000 dilution

Details

• Detection Notes: ECL, exposured for 3 min

reviewed by: Sonia Rocha ICC Human 12/21/2011

Summary

• Application: Immunocytochemistry
• Sample Tested: Human cells
• Species: Human
• Comments: Really nice results using microscopy.

Blocking

• Blocking Details: Blocking Buffer: 1% Donkey serum PBS, Blocking Time: 30 minutes, Blocking Temp: 20 degrees Celsius

Primary Anitbody

• Dilution Ratio: Incubation Dilution Buffer: PBS/1% Donkey serum, Dilution Ratio: 1:250, Incubation Time: 1 hour, Incubation Temp: 20 C

Secondary Antibody

• Secondary Description: Secondary Ab: anti-rabbit alexa 488, Secondary Ab Dilution: 1:500

Details

• Detection Notes: Detection Visualization Method: fluorescence at 488, Negative Control: secondary and primary only
• Fixation Details: Sample Preparation: PFA fixed

Comments

• Comments: Really nice results using microscopy.

FAQs for Carbonic Anhydrase IX/CA9 Antibody (NB100-417). (Showing 1 - 8 of 8 FAQs).

1. 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.
2. 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.
3. I'm going to use your CA-9 Antibody for paraffin-Immunohistochemistry. Do I have to do any special pretreatments?
   • Here is our protocol specific for this product. However, 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.
4. 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.
5. 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.
6. 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?
   • Our 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 the link here for those products: AbSelect.
7. 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.
8. 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.

Bioinformatics

• Gene Symbol: CA9
• Entrez:
  • 768(Human)
  • 230099(Mouse)