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Cardiovascular Blood Serum Proteins
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RecombinantSimpleStep

Human alpha Fetoprotein ELISA Kit (ab193765)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Human alpha Fetoprotein ELISA Kit (ab193765)
  • Human AFP standard curve comparison
  • Example of human AFP standard curve in Sample Diluent 75BS.
  • Interpolated concentrations of native AFP in pregnant human serum and HepG2 cell culture supernatant samples.
  • Serum from ten individual healthy human male donors was measured in duplicate.
  • Serum from ten individual healthy human female donors was measured in duplicate.
  • Sandwich ELISA - Human alpha Fetoprotein ELISA Kit (ab193765)

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

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Overview

  • Product name

    Human alpha Fetoprotein ELISA Kit
    See all alpha 1 Fetoprotein kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 2%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 4.5%
  • Sample type

    Cell culture supernatant, Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    44.8 pg/ml
  • Range

    156.25 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%
    Hep Plasma 96 94% - 97%
    EDTA Plasma 100 98% - 102%
    Cit plasma 89 89% - 89%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    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.

  • 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.

  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store 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 1 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
  • Research areas

    • Cardiovascular
    • Blood
    • Serum Proteins
    • Cancer
    • Tumor biomarkers
    • Other
    • Cancer
    • Tumor biomarkers
    • Tumor antigens
    • Developmental Biology
    • Organogenesis
    • Gut development
    • Liver development
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cardiovascular ELISA kits
  • 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.
  • Target information above from: UniProt accession P02771 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Afp
    • AFPD
    • Alpha fetoglobulin
    • Alpha fetoprotein
    • Alpha fetoprotein precursor
    • Alpha-1-fetoprotein
    • Alpha-fetoglobulin
    • Alpha-fetoprotein
    • alpha-fetoprotein, Hereditary persistence of, included
    • FETA
    • FETA_HUMAN
    • Hereditary persistence of alpha fetoprotein
    • HPAFP
    see all
  • Database links

    • Entrez Gene: 174 Human
    • Omim: 104150 Human
    • SwissProt: P02771 Human
    • Unigene: 518808 Human

    Associated products

    • Alternative Versions

      • Human alpha Fetoprotein Antibody Pair - BSA and Azide free (ab256602)

    Images

    • Other - Human alpha Fetoprotein ELISA Kit (ab193765)
      Other - Human alpha Fetoprotein ELISA Kit (ab193765)

      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.

       

    • Human AFP standard curve comparison
      Human AFP standard curve comparison

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

    • Example of human AFP standard curve in Sample Diluent 75BS.
      Example of human AFP standard curve in Sample Diluent 75BS.

      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.

    • Interpolated concentrations of native AFP in pregnant human serum and HepG2 cell culture supernatant samples.
      Interpolated concentrations of native AFP in pregnant human serum and HepG2 cell culture supernatant samples.

      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.

    • Serum from ten individual healthy human male donors was measured in duplicate.
      Serum from ten individual healthy human male donors was measured in duplicate.

      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.

    • Serum from ten individual healthy human female donors was measured in duplicate.
      Serum from ten individual healthy human female donors was measured in duplicate.

      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.

    • Sandwich ELISA - Human alpha Fetoprotein ELISA Kit (ab193765)
      Sandwich ELISA - Human alpha Fetoprotein ELISA Kit (ab193765)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (9)

    Publishing research using ab193765? Please let us know so that we can cite the reference in this datasheet.

    ab193765 has been referenced in 9 publications.

    • Al-Shahari EA  et al. Doxorubicin, L-arginine, or their combination as a prophylactic agent against hepatic carcinoma in mice. Environ Sci Pollut Res Int 28:37661-37671 (2021). PubMed: 33721166
    • 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

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