Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human Carbonic Anhydrase IX Quantikine ELISA Kit Summary
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.
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.
Precision
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 | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 166 | 338 | 673 | 166 | 333 | 647 |
| Standard Deviation | 6.36 | 7.19 | 20.5 | 10.5 | 21.6 | 39.7 |
| CV% | 3.8 | 2.1 | 3 | 6.3 | 6.5 | 6.1 |
Recovery
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 |
Linearity
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.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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; 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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- 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.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- 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.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- 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.
50 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human Carbonic Anhydrase IX Quantikine ELISA Kit
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.
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Citations: Showing 1 - 10
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Molecular profiling reveals a hypoxia signature in breast implant-associated anaplastic large cell lymphoma
Authors: N Oishi, T Hundal, JL Phillips, S Dasari, G Hu, DS Viswanatha, R He, M Mai, HK Jacobs, NH Ahmed, SI Syrbu, Y Salama, JR Chapman, F Vega, J Sidhu, NN Bennani, AL Epstein, LJ Medeiros, MW Clemens, RN Miranda, AL Feldman
Haematologica, 2021-06-01;0(0):.
Species: Human
Sample Types: Complex Sample Type
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Small cell lung cancer: model of circulating tumor cell tumorospheres in chemoresistance
Authors: L Klameth, B Rath, M Hochmaier, D Moser, M Redl, F Mungenast, K Gelles, E Ulsperger, R Zeillinger, G Hamilton
Sci Rep, 2017-07-13;7(1):5337.
Species: Human
Sample Types: Cell Culture Supernates
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DNA-linked Inhibitor Antibody Assay (DIANA) for sensitive and selective enzyme detection and inhibitor screening
Nucleic Acids Res, 2016-09-26;0(0):.
Species: Human
Sample Types: Cell Lysates
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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, 2016-01-30;14(1):31.
Species: Human
Sample Types: Urine
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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, 2012-10-19;7(10):e47469.
Species: Human
Sample Types: Urine
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Prognostic utility of pre-operative circulating osteopontin, carbonic anhydrase IX and CRP in renal cell carcinoma.
Br. J. Cancer, 2012-08-23;107(7):1131-7.
Species: Human
Sample Types: Plasma
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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., 2011-06-01;17(14):4872-4881.
Species: Human
Sample Types: Plasma
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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., 2011-03-02;48(0):112-20.
Species: Human
Sample Types: Serum
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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, 2010-05-11;102(11):1627-35.
Species: Human
Sample Types: Plasma
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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, 2008-10-07;99(9):1468-75.
Species: Human
Sample Types: Cell Culture Supernates
FAQs
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Reviews for Human Carbonic Anhydrase IX Quantikine ELISA Kit
Average Rating: 5 (Based on 2 Reviews)
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Human Carbonic Anhydrase IX Quantikine ELISA Kit
By Tanya Hundal
on 12/02/2020
Sample Tested: Human blood serum,human breast implant seroma,human breast implant-associated anaplastic large cell lymphoma,Human Plasma
Human Carbonic Anhydrase IX Quantikine ELISA Kit
By Aleksandra Gruslova
on 02/21/2018
Sample Tested: Human plasma samples
