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Recombinant Human Carbonic Anhydrase IX Protein, CF

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Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Carbonic Anhydrase IX Protein, CF Summary

Details of Functionality
Measured by its esterase activity. The specific activity is >10 pmol/min/µg, as measured with under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Carbonic Anhydrase IX/CA9 protein
Pro59-Asp414, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Pro59
Protein/Peptide Type
Recombinant Enzymes
Gene
CA9
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
42 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.
SDS-PAGE
53 kDa and 54 kDa, reducing conditions
Publications
Read Publications using
2188-CA in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5.
Assay Procedure
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase IX/CA9 (rhCA9) (Catalog # 2188-CA)
  • Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
  • 96-well Clear Plate (Costar, Catalog # 2592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhCA9 to 20 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load in plate, 50 µL of 20 ng/µL rhCA9, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
  4. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard 4-Nitrophenol (Sigma, Catalog # 241326).

Per Well:
  • rhCA9: 1 µg
  • Substrate: 1 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Carbonic Anhydrase IX Protein, CF

  • CA9
  • CAIX
  • CA-IX
  • Carbonate dehydratase IX
  • carbonic anhydrase 9
  • Carbonic Anhydrase IX
  • carbonic dehydratase
  • EC 4.2.1.1
  • G250
  • Membrane antigen MN
  • MN
  • P54/58N
  • PMW1
  • RCC
  • RCC-associated antigen G250
  • RCC-associated protein G250
  • Renal cell carcinoma-associated antigen G250

Background

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 (6th International Conference on the CAs, June 20‑25, 2003, 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). rhCA9 corresponds to the extracellular portion of human CA9.

  1. Pastorek, J. et al. (1994) Oncogene 9:2877.
  2. Opavsky, R. et al. (1996) Genomics 33:480.
  3. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  4. Kaluzova, M. et al. (2004) Mol. Cell Biol. 24:5757.
  5. Mukouyama, H. et al. (2004) Clin. Cancer Res. 10:1421.

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Recombinant Human
Carbonic Anhydrase IX ...
2188-CA
Recombinant Human Carbonic Anhydrase IX Protein, CF
Species: Hu
Applications: Enzyme Activity

Publications for Carbonic Anhydrase IX/CA9 (2188-CA)(4)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: Bioassay, ELISA (Control).


Filter By Application
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(2)
ELISA (Control)
(1)
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Showing Publications 1 - 4 of 4.
Publications using 2188-CA Applications Species
S Hao, X Cheng, X Wang, R An, H Xu, M Guo, C Li, Y Wang, Z Hou, C Guo Design, synthesis and biological evaluation of novel carbohydrate-based sulfonamide derivatives as antitumor agents Bioorg. Chem., 2020-08-27;104(0):104237. 2020-08-27 [PMID: 32911194] (Bioassay, Human) Bioassay Human
M Swayampaku, PC McDonald, M Vallejo, E Coyaud, SC Chafe, A Westerback, G Venkateswa, J Shankar, G Gao, EMN Laurent, Y Lou, KL Bennewith, CT Supuran, IR Nabi, B Raught, S Dedhar The interactome of metabolic enzyme carbonic anhydrase IX reveals novel roles in tumor cell migration and invadopodia/MMP14-mediated invasion Oncogene, 2017-07-10;36(45):6244-6261. 2017-07-10 [PMID: 28692057] (Bioassay) Bioassay
D Lemke, HW Pledl, M Zorn, M Jugold, E Green, J Blaes, S Löw, A Hertenstei, M Ott, F Sahm, AC Steffen, M Weiler, F Winkler, M Platten, Z Dong, W Wick Slowing down glioblastoma progression in mice by running or the anti-malarial drug dihydroartemisinin? Induction of oxidative stress in murine glioblastoma therapy Oncotarget, 2016-08-30;7(35):56713-56725. 2016-08-30 [PMID: 27447560]
Wind TC, Messenger MP, Thompson D, Selby PJ, Banks RE Measuring carbonic anhydrase IX as a hypoxia biomarker: differences in concentrations in serum and plasma using a commercial enzyme-linked immunosorbent assay due to influences of metal ions. Ann. Clin. Biochem., 2011-03-02;48(0):112-20. 2011-03-02 [PMID: 21367885] (ELISA (Control)) ELISA (Control)

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Understanding ‘Y’ in Breast Cancer: Crucial Role of DNA/RNA-binding Protein YB-1 in the Development, Pre-Invasive, and Metastatic Phases
Jamshed Arslan, Pharm D, PhD In the United States, 1 in 8 women will be diagnosed with breast cancer in her lifetime.1 Despite the prevalence, cancer genesis is a mystery. The heterogeneity of cancers makes it diff...  Read full blog post.
Carbonic anhydrase IX (CAIX) - a reliable histochemical marker of hypoxia
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Bioinformatics

Gene Symbol CA9
Uniprot