Human Carbonic Anhydrase IX/CA9 Antibody

Name: Human Carbonic Anhydrase IX/CA9 Antibody
Brand: R&D Systems
SKU: AF2188
Rating: 5 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF2188

Citations (7)

(1)

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Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase IX Pro59-Asp414 Accession # Q16790
Applications:	CyTOF-reported, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
Specificity:	Detects human Carbonic Anhydrase IX (CA9) in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse CA9 is observed and less than 1% cross-reactivity with recombinant human CA1, 3, 4, 8, 10, 12, and 14 is observed.

View Full Technical Details for Human Carbonic Anhydrase IX/CA9 Antibody

Applications for Human Carbonic Anhydrase IX/CA9 Antibody

	Recommended Concentration	Sample
Western Blot	1 µg/mL	See below
Flow Cytometry	0.25 µg/10⁶ cells	U87-MG cells
Immunohistochemistry	5-15 µg/mL	See below
Immunoprecipitation	25 µg/mL	Conditioned cell culture medium spiked with Recombinant Human Carbonic Anhydrase IX (Catalog # 2188-CA), see our available Western blot detection antibodies
CyTOF-reported	Chevrier, S. et al. (2018) Cell Systems. 6: 612. Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry	3-15 µg/mL	See below

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Human Carbonic Anhydrase IX/CA9 Antibody

	Western Blot Detection of Human Carbonic Anhydrase IX/CA9 by Western Blot. Western blot shows lysates of U-87 MG human glioblastoma/ astrocytoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Carbonic Anhydrase IX/CA9 at approximately 58 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
	Immunocytochemistry Carbonic Anhydrase IX/CA9 in A431 Human Cell Line. Carbonic Anhydrase IX/CA9 was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
	Immunohistochemistry Carbonic Anhydrase IX/CA9 in Human Colon Cancer Tissue. Carbonic Anhydrase IX/CA9 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
	Flow Cytometry Detection of Carbonic Anhydrase IX/CA9 in U87-MG cells cells by Flow Cytometry. U87-MG cells were stained with Goat Anti-Human Carbonic Anhydrase IX/CA9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2188, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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Background: Carbonic Anhydrase IX/CA9

Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO₂ + H₂O = HCO₃^- + H⁺, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (1-3). Topics in the CA meeting (6^th International Conference on the CAs, June 20-25, 2003, in Slovakia) ranged from use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO₂ removal and mosquito control in industry. CA9, also known as membrane antigen MN and renal cell carcinoma (RCC)-associated antigen G250, is a transmembrane enzyme expressed primarily in carcinoma cells. It is one of the best markers for hypoxia and for RCC (4, 5). Recombinant human CA9 corresponds to the extracellular portion of human CA9.

Item	Value
References	Pastorek, J. et al. (1994) Oncogene 9: 2877. Opavsky, R. et al. (1996) Genomics 33: 480. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50. Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757. Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.
Entrez Gene IDs	768 (Human); 230099 (Mouse)
Alternate Names	CA-IX, CA9, CAIX, Carbonate dehydratase IX, Carbonic Anhydrase IX, EC 4.2.1.1, G250, MN, Membrane antigen MN, P54/58N, PMW1, RCC, RCC-associated antigen G250, RCC-associated protein G250, Renal cell carcinoma-associated antigen G250, carbonic anhydrase 9, carbonic dehydratase

Citations for Human Carbonic Anhydrase IX/CA9 Antibody (7)

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Citations are publications that use Bio-Techne products. Selected citations for Human Carbonic Anhydrase IX/CA9 Antibody include:

Species: Human
Sample Types: Whole Tissue
Applications: IHC

S Lefort et al. (2021), De novo and cell line models of human mammary cell transformation reveal an essential role for Yb-1 in multiple stages of human breast cancer Cell Death and Differentiation, 0(0).
PMID: 34294889
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

SC Chafe et al. (2021), Genome-wide synthetic lethal screen unveils novel CAIX-NFS1/xCT axis as a targetable vulnerability in hypoxic solid tumors Science Advances, 7(35).
PMID: 34452919
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot

EJ Campbell et al. (2019), Activation of the hypoxia pathway in breast cancer tissue and patient survival are inversely associated with tumor ascorbate levels BMC Cancer, 19(1):307.
PMID: 30943919
Species: Xenograft
Sample Types: Whole Tissue
Applications: IHC

EE Hedlund et al. (2019), Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts Cancers (Basel), 11(7).
PMID: 31319613
Species: Human
Sample Types: Whole Cells
Applications: CyTof

Chevrier S et al. (2018), Compensation of Signal Spillover in Suspension and Imaging Mass Cytometry. Cell Syst, 0(0).
PMID: 29605184
Species: Human
Sample Types: Whole Tissue
Applications: CyTof

S Chevrier et al. (2018), Compensation of Signal Spillover in Suspension and Imaging Mass Cytometry Cell Syst, 0(0).
PMID: 29605184
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

M Swayampaku et al. (2017), The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion Oncogene, 36(45):6244-6261.
PMID: 28692057

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Human Carbonic Anhydrase IX/CA9 Antibody

Name: Santhosh Sivajothi

Application: IHC

Sample Tested: Renal cancer tissue

Species: Human

Verified Customer | Posted 05/21/2018

Immunohistochemistry was performed on FFPE tissue section. Heat induced antigen retrieval was performed by heating section in citrate buffer (pH 6) at 95C for 20 minutes.

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Human Carbonic Anhydrase IX/CA9 Antibody

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Applications for Human Carbonic Anhydrase IX/CA9 Antibody

Scientific Data Examples for Human Carbonic Anhydrase IX/CA9 Antibody

Western Blot

Immunocytochemistry

Immunohistochemistry

Flow Cytometry

Formulation, Preparation and Storage

Reconstitution Calculator

Background: Carbonic Anhydrase IX/CA9

Citations for Human Carbonic Anhydrase IX/CA9 Antibody (7)

Customer Reviews for Human Carbonic Anhydrase IX/CA9 Antibody (1)

Customer Images

Human Carbonic Anhydrase IX/CA9 Antibody

Product Documents for Human Carbonic Anhydrase IX/CA9 Antibody

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