Overview

  • Product name
    Human alpha Fetoprotein ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 8 10%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 6.9%
  • Sample type
    Cell culture supernatant, Urine, Serum, Cell culture media, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    65.1 pg/ml
  • Range
    156 pg/ml - 10000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 103 101% - 113%
    Urine 119 116% - 121%
    Serum 91 88% - 96%
    Cell culture media 94 91% - 98%
    Heparin Plasma 96 94% - 97%
    EDTA Plasma 100 98% - 102%
    Citrate Plasma 89 89% - 89%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse
  • Product overview

    Abcam’s alpha Fetoprotein in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of alpha Fetoprotein protein in human serum, plasma and cell culture supernatant.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:

    Samples diluted in Sample Diluent NS – 87 pg/mL
    Samples diluted in Sample Diluent 25BS – 178 pg/mL

  • Notes

    Alpha-fetoprotein (AFP) is a secreted single chain glycoprotein expressed normally during fetal development as well as by liver pathologies, including cancer. It is a member of the albuminoid superfamily and may act as a carrier protein. Pregnant women may have elevated alpha-fetoprotein levels derived from fetal circulation and male and female adults have low levels of circulating alpha-fetoprotein.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human alpha Fetoprotein Capture Antibody 1 x 600µl
    10X Human alpha Fetoprotein Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Human alpha Fetoprotein Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 5BI 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent 25BS 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Function
    Binds copper, nickel, and fatty acids as well as, and bilirubin less well than, serum albumin. Only a small percentage (less than 2%) of the human AFP shows estrogen-binding properties.
  • Tissue specificity
    Plasma. Synthesized by the fetal liver and yolk sac.
  • Sequence similarities
    Belongs to the ALB/AFP/VDB family.
    Contains 3 albumin domains.
  • Developmental stage
    Occurs in the plasma of fetuses more than 4 weeks old, reaches the highest levels during the 12th-16th week of gestation, and drops to trace amounts after birth. The serum level in adults is usually less than 40 ng/ml. AFP occurs also at high levels in the plasma and ascitic fluid of adults with hepatoma.
  • Post-translational
    modifications
    Independent studies suggest heterogeneity of the N-terminal sequence of the mature protein and of the cleavage site of the signal sequence.
    Sulfated.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • Afp
    • Alpha-1-fetoprotein
    • Alpha-fetoglobulin
    • Alpha-fetoprotein
    • FETA
    • FETA_HUMAN
    • HPAFP
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab193765 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Standard Curve comparison between human AFP SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • The AFP standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of AFP were measured in duplicates, interpolated from the AFP standard curves and corrected for sample dilution. Undiluted samples are as follows: pregnant human serum 3% and HepG2 cell culture supernatant 1:666. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 221.8 ng/mL in neat pregnant human serum and 6,165 ng/mL in neat HepG2 cell culture supernatant.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration was determined to be 678.4 pg/mL with a range of 133.9 – 2,257 pg/mL.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AFP concentration in nine detectable donor samples was determined to be 550.7 pg/mL with a range of 247.9-816.1 pg/mL. Donor #5 was not detectable.

Protocols

References

This product has been referenced in:
  • Chen D  et al. Activated Wnt signaling promotes growth and progression of AFP-producing gastric cancer in preclinical models. Cancer Manag Res 11:1349-1362 (2019). Read more (PubMed: 30809100) »
  • Wang X  et al. Extracellular mRNA detected by molecular beacons in tethered lipoplex nanoparticles for diagnosis of human hepatocellular carcinoma. PLoS One 13:e0198552 (2018). Read more (PubMed: 29879168) »
See all 4 Publications for this product

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