Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Related Research Areas
Human HE4/WFDC2 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 (50 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL), Saliva (10 uL), Urine (10 uL)
Sensitivity
32.2 pg/mL
Assay Range
78.1 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human HE4/WFDC2
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Human HE4/WFDC2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human HE4/WFDC2 in cell culture supernates, serum, plasma, saliva, and urine. It contains NS0-expressed recombinant human HE4/WFDC2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human HE4/WFDC2 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 HE4/WFDC2.
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, Saliva
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 470 | 1278 | 2448 | 476 | 1408 | 2731 |
| Standard Deviation | 11.5 | 35.3 | 86.1 | 26 | 79.1 | 162 |
| CV% | 2.4 | 2.8 | 3.5 | 5.5 | 5.6 | 5.9 |
Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 451 | 1337 | 2629 | 452 | 1409 | 2761 |
| Standard Deviation | 14 | 39.8 | 85.7 | 25 | 78 | 202 |
| CV% | 3.1 | 3 | 3.3 | 5.5 | 5.5 | 7.3 |
Recovery
The recovery of HE4/WFDC2 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=8) | 103 | 95-114 |
| EDTA Plasma (n=4) | 108 | 94-115 |
| Heparin Plasma (n=4) | 107 | 96-115 |
| Saliva (n=4) | 100 | 98-107 |
| Serum (n=4) | 104 | 92-115 |
| Urine (n=4) | 98 | 85-111 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of HE4/WFDC2 were serially diluted with 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.
⚠ 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 the appropriate Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- 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.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. 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
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human HE4/WFDC2 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 - 8
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Diagnostic value of circN4BP2L2 in type I and type II epithelial ovarian cancer
Authors: L Ning, J Lang, B Long, L Wu
BMC Cancer, 2022-11-24;22(1):1210.
Species: Human
Sample Types: Plasma
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Plasma circN4BP2L2 is a promising novel diagnostic biomarker for epithelial ovarian cancer
Authors: L Ning, J Lang, L Wu
BMC Cancer, 2022-01-03;22(1):6.
Species: Human
Sample Types: Serum
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Serum human epididymis protein 4 level as a predictor of clinical worsening in idiopathic pulmonary arterial hypertension: a pilot study
Authors: Q Jin, Y Tang, Z Liu, W Xie, Q Luo, Z Zhao, Q Zhao, Z Huang, X Yu, L Yan, C Xiong, X Ni, Y Yan
BMC Cardiovasc Disord, 2020-04-15;20(1):175.
Species: Human
Sample Types: Serum
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Development of Potent Forchlorfenuron Analogs and Their Cytotoxic Effect in Cancer Cell Lines
Authors: KK Kim, RK Singh, N Khazan, A Kodza, NA Singh, A Jones, U Sivagnanal, M Towner, H Itamochi, R Turner, RG Moore
Sci Rep, 2020-02-24;10(1):3241.
Species: Human
Sample Types: Cells
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Colonic epithelial cell diversity in health and inflammatory bowel disease
Authors: K Parikh, A Antanavici, D Fawkner-Co, M Jagielowic, A Aulicino, C Lagerholm, S Davis, J Kinchen, HH Chen, NK Alham, N Ashley, E Johnson, P Hublitz, L Bao, J Lukomska, RS Andev, E Björklund, BM Kessler, R Fischer, R Goldin, H Koohy, A Simmons
Nature, 2019-02-27;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Noninvasive ovarian cancer biomarker detection via an optical nanosensor implant
Authors: RM Williams, C Lee, TV Galassi, JD Harvey, R Leicher, M Sirenko, MA Dorso, J Shah, N Olvera, F Dao, DA Levine, DA Heller
Sci Adv, 2018-04-18;4(4):eaaq1090.
Species: Human, Mouse
Sample Types: Ascites Fluid, Serum
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Expression and diagnostic value of HE4 in pancreatic adenocarcinoma.
Authors: Huang, Tianhe, Jiang, Shi-Wen, Qin, Liangyi, Senkowski, Christop, Lyle, Christia, Terry, Karen, Brower, Steven, Chen, Haibin, Glasgow, Wayne, Wei, Yongchan, Li, Jinping
Int J Mol Sci, 2015-01-29;16(2):2956-70.
Species: Human
Sample Types: Plasma
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Kallikrein family proteases KLK6 and KLK7 are potential early detection and diagnostic biomarkers for serous and papillary serous ovarian cancer subtypes.
Authors: Tamir, Ayala, Jag, Ushma, Sarojini, Sreeja, Schindewolf, Craig, Tanaka, Takemi, Gharbaran, Rajendra, Patel, Hiren, Sood, Anil, Hu, Wei, Patwa, Ruzeen, Blake, Patrick, Chirina, Polina, Oh Jeong, Jin, Lim, Heejin, Goy, Andre, Pecora, Andrew, Suh, K Stephe
J Ovarian Res, 2014-12-05;7(1):109.
Species: Human
Sample Types: Serum
FAQs
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