Human HE4/WFDC2 Quantikine ELISA Kit

  (3 citations)     
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Assay Procedure
Citations (3)
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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 (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.
Control Available
QC102 , Quantikine Immunoassay Control Set 742 for Human HE4/WFDC2 - Please Inquire
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 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 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.
 HE4/WFDC2 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: HE4/WFDC2
  • Long Name:
    WAP Four-disulfide Core Domain protein 2
  • Entrez Gene IDs:
    10406 (Human); 67701 (Mouse); 286888 (Rat)
  • Alternate Names:
    dJ461P17.6; EDDM4; Epididymal secretory protein E4; epididymis-specific, whey-acidic protein type, four-disulfide core; HE4; HE4MGC57529; Major epididymis-specific protein E4; Putative protease inhibitor WAP5; WAP domain containing protein HE4-V4; WAP four-disulfide core domain 2; WAP four-disulfide core domain protein 2; WAP5; WAP5epididymal protein 4; WFDC2
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 the appropriate Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  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.
  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. 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.
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Sample Type
  1. 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;4(4):eaaq1090.
    Species: Human
    Sample Type: Serum
  2. 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;16(2):2956-70.
    Species: Human
    Sample Type: Plasma
  3. 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;7(1):109.
    Species: Human
    Sample Type: Serum

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ELISA Controls
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Quantikine Immunoassay Control Set 742 for Human HE4/WFDC2

Ctrl QC102
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Supplemental ELISA Products
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Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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