Human Carbonic Anhydrase IX Quantikine ELISA Kit

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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL), Urine (100 uL)
  • Sensitivity
    4.39 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human CA9
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
Product Summary
The Quantikine Human Carbonic Anhydrase IX Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CA9 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human CA9 ectodomain and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CA9 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human CA9.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Standard Deviation6.367.1920.510.521.639.7


The recovery of CA9 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 110 103-114
Citrate Plasma (n=4) 96 89-111
EDTA Plasma (n=4) 99 90-110
Heparin Plasma (n=4) 97 88-112
Serum (n=4) 97 87-106
Urine (n=4) 101 89-114
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of CA9 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Carbonic Anhydrase IX Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Carbonic Anhydrase IX/CA9
Carbonic anhydrases catalyze the reversible reaction of CO2 + H2O = HCO3- + H+, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption. Human members of the CAs are secreted (CA6), membrane-bound (CA4, 9 to 12 and 14), cytosolic (CA1 to 3, 7, 8 and 13) or mitochondrial (CA5A and 5B). Some CAs may serve as markers for tumors and hypoxia. A subset of CAs lack CA activity due to point mutations but retain esterase function.
    • Entrez Gene IDs
      768 (Human); 230099 (Mouse);
    • Alternate Names
      CA9; CA-IX; CAIXcarbonic anhydrase 9; Carbonate dehydratase IX; carbonic anhydrase IXpMW1; carbonic dehydratase; EC; G250; Membrane antigen MN; MN; MNRCC-associated antigen G250; P54/58N; RCC; RCC-associated protein G250; Renal cell carcinoma-associated antigen G250;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    Showing Results 1 - 7 of 7
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    Sample Type
    1. A multiplex immunoassay for the non-invasive detection of bladder cancer.
      Authors: Shimizu Y, Furuya H, Bryant Greenwood P, Chan O, Dai Y, Thornquist M, Goodison S, Rosser C
      J Transl Med, 2016;14(1):31.
      Species: Human
      Sample Type: Urine
    2. A multi-analyte assay for the non-invasive detection of bladder cancer.
      Authors: Goodison, Steve, Chang, Myron, Dai, Yunfeng, Urquidi, Virginia, Rosser, Charles
      PLoS ONE, 2012;7(10):e47469.
      Species: Human
      Sample Type: Urine
    3. Prognostic utility of pre-operative circulating osteopontin, carbonic anhydrase IX and CRP in renal cell carcinoma.
      Br. J. Cancer, 2012;107(7):1131-7.
      Species: Human
      Sample Type: Plasma
    4. Myeloid Biomarkers Associated with Glioblastoma Response to Anti-VEGF Therapy with Aflibercept.
      Authors: de Groot JF, Piao Y, Tran H, Gilbert M, Wu HK, Liu J, Bekele BN, Cloughesy T, Mehta M, Robins HI, Lassman A, Deangelis L, Camphausen K, Chen A, Yung W, Prados M, Wen PY, Heymach JV
      Clin. Cancer Res., 2011;17(14):4872-4881.
      Species: Human
      Sample Type: Plasma
    5. 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.
      Authors: Wind TC, Messenger MP, Thompson D, Selby PJ, Banks RE
      Ann. Clin. Biochem., 2011;48(0):112-20.
      Species: Human
      Sample Type: Serum
    6. High levels of carbonic anhydrase IX in tumour tissue and plasma are biomarkers of poor prognostic in patients with non-small cell lung cancer.
      Authors: Ilie M, Mazure NM, Hofman V, Ammadi RE, Ortholan C, Bonnetaud C, Havet K, Venissac N, Mograbi B, Mouroux J, Pouyssegur J, Hofman P
      Br. J. Cancer, 2010;102(11):1627-35.
      Species: Human
      Sample Type: Plasma
    7. Expression of carbonic anhydrase 9, a potential intrinsic marker of hypoxia, is associated with poor prognosis in oesophageal squamous cell carcinoma.
      Authors: Tanaka N, Kato H, Inose T, Kimura H, Faried A, Sohda M, Nakajima M, Fukai Y, Miyazaki T, Masuda N, Fukuchi M, Kuwano H
      Br. J. Cancer, 2008;99(9):1468-75.
      Species: Human
      Sample Type: Cell Culture Supernates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Group 9

    Ctrl QC24
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    Supplemental ELISA Products
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    Quantikine Wash Buffer 1

    ELISA WA126 5
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