Human CA125/MUC16 Quantikine ELISA Kit

(2 citations)   
  • 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 (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (10 uL), Urine (100 uL), Human Milk (10 uL)
  • Sensitivity
    0.099 U/mL
  • Assay Range
    0.5 - 32 U/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
  • Specificity
    Natural and recombinant human CA125
  • 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 CA125/MUC16 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human CA125 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains CHO cell-expressed recombinant human CA125 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human CA125 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 CA125.

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, Urine, Human Milk
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.0660.1010.2660.3130.5270.666


The recovery of CA125 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 95-102
EDTA Plasma (n=4) 97 93-101
Heparin Plasma (n=4) 97 92-100
Serum (n=4) 97 90-101
To assess the linearity of the assay, samples containing high concentrations of CA125 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CA125/MUC16 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CA125/MUC16
The CA125 antigen, also known as MUC16, is a mucin protein that may be found in type I transmembrane or secreted forms that are used monitor the progress of epithelial ovarian cancer therapy. Expression of isoforms, proteolytic cleavage, and heavy N- and O- linked glycosylation produce forms of human CA125 that can vary from 200-5000 kDa in size. It inhibits NK cell activity, which is thought to contribute to immune evasion in peritoneal cancer and pregnancy.
    • Long Name
      Ovarian Cancer Antigen CA125
    • Entrez Gene IDs
      94025 (Human);
    • Alternate Names
      CA125 ovarian cancer antigen; CA-125; CA125MUC-16; FLJ14303; MUC16; mucin 16, cell surface associated; mucin-16; Ovarian cancer-related tumor marker CA125; Ovarian carcinoma antigen CA125;
    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. 100 µL Assay Diluent
    4.   Add 100 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.

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    Sample Type
    1. 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
    2. False biomarker discovery due to reactivity of a commercial ELISA for CUZD1 with cancer antigen CA125.
      Authors: Prassas, Ioannis, Brinc, Davor, Farkona, Sofia, Leung, Felix, Dimitromanolakis, Apostolo, Chrystoja, Caitlin, Brand, Randall, Kulasingam, Vathany, Blasutig, Ivan M, Diamandis, Elefther
      Clin Chem, 2014;60(2):381-8.
      Species: Human
      Sample Type: Serum
      Application: WB
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