Key features and details
- One-wash 90 minute protocol
- Sensitivity: 44.8 pg/ml
- Range: 156.25 pg/ml - 10000 pg/ml
- Sample type: Cell culture media, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman alpha Fetoprotein ELISA Kit
See all alpha 1 Fetoprotein kits
Intra-assay Sample n Mean SD CV% Serum 8 2% Inter-assay Sample n Mean SD CV% Serum 3 4.5%
Sample typeCell culture supernatant, Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma
Assay typeSandwich (quantitative)
Range156.25 pg/ml - 10000 pg/ml
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% Hep Plasma 96 94% - 97% EDTA Plasma 100 98% - 102% Cit plasma 89 89% - 89%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Human alpha Fetoprotein ELISA Kit (ab193765) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of alpha Fetoprotein protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, urine, and cell culture media. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human alpha Fetoprotein with 44.8 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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.
PlatformMicroplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 5BI 1 x 6ml Human alpha Fetoprotein Capture Antibody (Lyophilized) 1 vial Human alpha Fetoprotein Detector Antibody (Lyophilized) 1 vial Human alpha Fetoprotein Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent 75BS 2 x 20ml Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
FunctionBinds 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 specificityPlasma. Synthesized by the fetal liver and yolk sac.
Sequence similaritiesBelongs to the ALB/AFP/VDB family.
Contains 3 albumin domains.
Developmental stageOccurs 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.
modificationsIndependent studies suggest heterogeneity of the N-terminal sequence of the mature protein and of the cleavage site of the signal sequence.
- Information by UniProt
- Alpha fetoglobulin
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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.
To learn more about the advantages of recombinant antibodies see here.
ab193765 has been referenced in 8 publications.
- Lacombe M et al. Comprehensive and comparative exploration of the Atp7b-/- mouse plasma proteome. Metallomics 12:249-258 (2020). PubMed: 31815268
- 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). PubMed: 30809100
- Pettinato G et al. Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells. Sci Rep 9:8920 (2019). PubMed: 31222080
- Zhang B et al. Prussian blue nanoparticle-labeled aptasensing platform on graphene oxide for voltammetric detection of a-fetoprotein in hepatocellular carcinoma with target recycling. Analyst 144:4858-4864 (2019). PubMed: 31294738
- Lv S et al. H2-Based Electrochemical Biosensor with Pd Nanowires@ZIF-67 Molecular Sieve Bilayered Sensing Interface for Immunoassay. Anal Chem 91:12055-12062 (2019). PubMed: 31436433
- 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). PubMed: 29879168
- Pettinato G et al. Scalable Differentiation of Human iPSCs in a Multicellular Spheroid-based 3D Culture into Hepatocyte-like Cells through Direct Wnt/ß-catenin Pathway Inhibition. Sci Rep 6:32888 (2016). Sandwich ELISA ; Human . PubMed: 27616299
- Cui Y et al. Biofunctionalized dendritic polyaniline nanofibers for sensitive electrochemical immunoassay of biomarkers. Analyst 137:1656-62 (2012). ELISA, Sandwich ELISA ; Human . PubMed: 22355804