Human alpha-Fetoprotein Quantikine ELISA Kit

(6 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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    0.046 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural human AFP
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    Interference observed with other substances.
Control Available
Product Summary
The Quantikine Human AFP immunoassay is a 4.5 hour solid phase ELISA designed to measure human AFP in cell culture supernates, serum, and plasma.

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
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.1090.2190.4410.1580.2610.549

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.080.1490.4470.2150.3820.93


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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 95-103
EDTA Plasma (n=4) 104 94-114
Heparin Plasma (n=4) 103 95-115
Serum (n=4) 103 86-115
To assess the linearity of the assay, samples spiked with high concentrations of AFP were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human alpha-Fetoprotein Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: alpha-Fetoprotein/AFP
alpha-Fetoprotein (AFP), is albuminoid superfamily protein that is synthesized in the fetus primarily by the liver, yolk sac, and tissues of gastrointestinal origin. It is one of the earliest markers of the hepatocyte lineage. AFP acts as a carrier protein for steroids, bilirubin, fatty acids, retinoids, and flavonoids. In addition, it can exert immunosuppressive activity, regulate cell proliferation and apoptosis, initiate intracellular signaling, and contribute to cell invasion. Altered levels of both fetal and maternal AFP have been associated with hypothyroidism, autoimmune disorders, and heart defects. Low maternal serum AFP levels are associated with a higher incidence of Down syndrome, whereas higher levels are associated with spina bifida and anencephaly. Elevated AFP levels are also coincident with liver, stomach, and germ cell cancers.
    • Entrez Gene IDs
      174 (Human); 11576 (Mouse); 24177 (Rat);
    • Alternate Names
      AFP; Alpha-1-fetoprotein; alpha-fetoglobulin; alpha-fetoprotein; DSCAM2; FETA; HP; HPAFP;
    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. 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.

    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 - 6 of 6
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    Sample Type
    1. Knockdown of beta-catenin with dicer-substrate siRNAs reduces liver tumor burden in vivo.
      Authors: Dudek H, Wong D, Arvan R, Shah A, Wortham K, Ying B, Diwanji R, Zhou W, Holmes B, Yang H, Cyr W, Zhou Y, Shah A, Farkiwala R, Lee M, Li Y, Rettig G, Collingwood M, Basu S, Behlke M, Brown B
      Mol Ther, 2014;22(1):92-101.
      Species: Human
      Sample Type: Serum
    2. Proteomics mapping of cord blood identifies haptoglobin "switch-on" pattern as biomarker of early-onset neonatal sepsis in preterm newborns.
      Authors: Buhimschi CS, Bhandari V, Dulay AT, Nayeri UA, Abdel-Razeq SS, Pettker CM, Thung S, Zhao G, Han YW, Bizzarro M, Buhimschi IA
      PLoS ONE, 2011;6(10):e26111.
      Species: Human
      Sample Type: Serum
    3. DLK1, a serum marker for hepatoblastoma in young infants.
      Authors: Falix FA, Aronson DC, Lamers WH, Hiralall JK, Seppen J
      Pediatr Blood Cancer, 2011;59(0):743.
      Species: Human
      Sample Type: Serum
    4. A hypoxia- and alpha-fetoprotein-dependent oncolytic adenovirus exhibits specific killing of hepatocellular carcinomas.
      Authors: Kwon OJ, Kim PH, Huyn S, Wu L, Kim M, Yun CO
      Clin. Cancer Res., 2010;16(24):6071-82.
      Species: Mouse
      Sample Type: Serum
    5. Identifying serological biomarkers of hepatocellular carcinoma using surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy.
      Authors: Wu FX, Wang Q, Zhang ZM, Huang S, Yuan WP, Liu JY, Ban KC, Zhao YN
      Cancer Lett., 2009;279(2):163-70.
      Species: Human
      Sample Type: Serum
    6. A novel approach and protocol for discovering extremely low-abundance proteins in serum.
      Authors: Tanaka Y, Akiyama H, Kuroda T, Jung G, Tanahashi K, Sugaya H, Utsumi J, Kawasaki H, Hirano H
      Proteomics, 2006;6(17):4845-55.
      Species: Human
      Sample Type: Serum
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 566 Human a-Fetoprotein

    Ctrl QC38
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine Wash Buffer 1

    ELISA WA126 5
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