antibody

Carbonic Anhydrase IX (CA9) (PE)

MBS603443

100 Tests

695 USD

monoclonal

mouse

PE

[7H20]

flow cytometry, FACS

Antibody

Carbonic Anhydrase IX (CA9) (PE)

[Carbonic Anhydrase IX]

[Anti -Carbonic Anhydrase IX (CA9) (PE)]

[carbonic anhydrase IX; Carbonic anhydrase 9; carbonic anhydrase 9; pMW1; CA-IX; P54/58N; OTTHUMP00000022773; membrane antigen MN; carbonic dehydratase; carbonate dehydratase IX; RCC-associated antigen G250; RCC-associated protein G250; renal cell carcinoma-associated antigen G250; carbonic anhydrase IX; Carbonate dehydratase IX; Carbonic anhydrase IX; CA-IX; CAIX; Membrane antigen MN; P54/58N; Renal cell carcinoma-associated antigen G250; RCC-associated antigen G250; pMW1]

[CA9; CA9; MN; CAIX; G250; MN]

CAH9_HUMAN

Monoclonal

IgG2a

[7H20]

mouse

Human

Recognizes human Carbonic Anhydrase IX (CA9). Does not crossreact with recombinant mouse CA9 or with recombinant human CA1, 2, 3, 4, 5A, 6, 7, 8, 10, 12, 13, or 14.

Affinity Purified. Purified by Protein A/G affinity chromatography.

Supplied as a liquid in saline, 0.5% BSA, 0.09% sodium azide. Labeled with R-Phycoerythrin (PE).

0.025mg/ml

May be stored at 4 degree C before opening. DO NOT FREEZE! Stable at 4 degree C as an undiluted liquid. Stable for 12 months at 4 degree C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.

Flow Cytometry (FC/FACS)

Suitable for use in Flow Cytometry. Dilution: Flow Cytometry: Neat. 10ul labels 1x10e5 cells. Tested using U-87 MG human glioblastoma/astrocytoma cell line. Optimal dilutions to be dertermined by the researcher.

Flow Cytometry (FC/FACS)

Immunogen: Recombinant corresponding to aa59-414 of human Carbonic Anhydrase IX (CA9) expressed in NSO cells (Q16790).

Flow Cytometry Protocol: Reagents Not Provided: . -PBS (Dulbecco's PBS) -BSA . Intended Use: . Designed to quantitatively determine the percentage of cells bearing Carbonic Anhydrase IX (CA9) within a population and qualitatively determine the density of CA9 on cell surfaces by flow cytometry. Principle of the Test: . Washed cells are incubated with MBS603443, which binds to cells expressing CA9. Unbound phycoerythrin-conjugated antibody is then washed from the cells. Cells expressing CA9 are fluorescently stained: with the intensity of staining directly proportional to the density of expression of CA9. Cell surface expression of CA9 is determined by flow cytometric analysis using 488nm wavelength laser excitation and monitoring emitted fluorescence with a detector optimized to collect peak emissions at 565-605nm. Sample Preparation: . Peripheral Blood Cells:. Whole blood should be collected in evacuated tubes containing EDTA or heparin as the anticoagulant. Contaminating serum components should be removed by washing the cells three times in an isotonic phosphate buffer (supplemented With 0.50/0 BSA) by centrifugation at 500 x g for 5 minutes. Transfer 5Oul of packed cells to a 5ml tube for staining with the monoclonal antibody. Whole blood will require lysis of RBC following the staining procedure. Cell Cultures: Continuous cell lines or activated cell cultures should be centrifuged at 500 x g for 5 minutes and washed three times in an isotonic PBS buffer (supplemented with 0.50/0 BSA), as described above, to remove any residual growth factors that may be present in the culture medium. Cells should then be resuspended in the same buffer to a final concentration of 4xl Oe6 cells/ml and 25ul of cells (1 xl Oe5) transferred to a Sml tube for staining. Note: Adherent cell lines may require pretreatment with O.5mM EDTA to facilitate removal from substrate. Cells that require trypsinization to enable removal from substrate should be further incubated in medium for 6-10 hours on a rocker platform to enable regeneration of the receptors. The use of the rocker platform will prevent reattachment to the substrate. Sample Staining: . 1. Cells should be Fe-blocked by treatment with 1 ug of human IgG/ lOe5 cells for 15 minutes at RT prior to staining. Do not wash excess blocking IgG from this reaction. 2. Transfer 25ul of the Fe-blocked cells (1 xl Oe5 cells) or SOul of packed whole blood to a 5ml tube. 3. Add 1 Oul of Cll 05-80A. Incubate for 30-45 minutes at 4°C. 4. Following this incubation, remove unreacted CA9 reagent by washing the cells twice in 4ml of the same PBS buffer (note: whole blood will require an RBC lysis step at this point using any commercially available lysing reagent). 5. Finally, resuspend the cells in 200-400ul of PBS buffer for final flow cytometric analysis. 6. As a control for analysis, cells in a separate tube should be treated with PE-Iabeled mouse IgG2a antibody. Optimai conditions should be determined by each laboratory.

Antibodies; Abs to Enzymes

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. 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.

119578763

Q16790

HIF-1-alpha Transcription Factor Network Pathway (138045); Nitrogen Metabolism Pathway (83025); Nitrogen Metabolism Pathway (416)

Carbonic anhydrases (CAs) are a large family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide. They participate in a variety of biological processes, including respiration, calcification, acid-base balance, bone resorption, and the formation of aqueous humor, cerebrospinal fluid, saliva, and gastric acid. They show extensive diversity in tissue distribution and in their subcellular localization. CA IX is a transmembrane protein and the only tumor-associated carbonic anhydrase isoenzyme known. It is expressed in all clear-cell renal cell carcinoma, but is not detected in normal kidney or most other normal tissues. It may be involved in cell proliferation and transformation. This gene was mapped to 17q21.2 by fluorescence in situ hybridization, however, radiation hybrid mapping localized it to 9p13-p12. [provided by RefSeq]

CA9: Reversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia. Forms oligomers linked by disulfide bonds. By hypoxia. Expressed primarily in carcinoma cells lines. Expression is restricted to very few normal tissues and the most abundant expression is found in the epithelial cells of gastric mucosa. Inhibited by coumarins, saccharin, sulfonamide derivatives such as acetazolamide (AZA) and Foscarnet (phosphonoformate trisodium salt). Belongs to the alpha-carbonic anhydrase family. Protein type: Membrane protein, integral; Energy Metabolism - nitrogen; Lyase; Nucleolus; EC 4.2.1.1. Chromosomal Location of Human Ortholog: 9p13.3. Cellular Component: basolateral plasma membrane; microvillus membrane; nucleolus; plasma membrane; integral to membrane. Molecular Function: carbonate dehydratase activity; zinc ion binding. Biological Process: response to drug; secretion; bicarbonate transport; morphogenesis of an epithelium; one-carbon compound metabolic process; response to testosterone stimulus

100 Tests

695 USD