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    products/elisa/human-transferrin-elisa-kit-ab187391.pdf

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SimpleStep

Human Transferrin ELISA Kit (ab187391)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (2)References (4)

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Other - Human Transferrin ELISA Kit (ab187391)
  • Human Transferrin standard curve comparison
  • Example of human Transferrin standard curve in Sample Diluent NS.
  • Standard curve
  • Interpolated concentrations of native Transferrin in human serum and plasmas samples.
  • Serum from seven individual healthy human female donors was measured in duplicate.
  • Linearity of dilution.
  • Sandwich ELISA - Human Transferrin ELISA Kit (ab187391)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 0.39 ng/ml
  • Range: 0.63 ng/ml - 40 ng/ml
  • Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human Transferrin ELISA Kit
    See all Transferrin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 5.2%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 8.3%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    0.39 ng/ml
  • Range

    0.63 ng/ml - 40 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 92 86% - 104%
    Cell culture media 101 97% - 103%
    Hep Plasma 114 112% - 116%
    EDTA Plasma 107 95% - 115%
    Cit plasma 115 104% - 121%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human Transferrin ELISA Kit (ab187391) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Transferrin protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Transferrin with 0.39 ng/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.


    ASSAY SPECIFICITY  This kit recognizes both native and recombinant human Transferrin protein in serum, plasma, and cell culture supernatant samples only. Urine, saliva, and cell/tissue extract samples have not been tested with this kit.


    CROSS REACTIVITY Human albumin, ferritin, and hemoglobin were prepared at 40 ng/mL and assayed for cross reactivity. No cross-reactivity was observed.


    SPECIES REACTIVITY This kit recognizes human Transferrin protein.
    Other species reactivity was determined by measuring 1:50,000 serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution. Reactivity < 3% was determined for the following species: Mouse, Rat, Cow

  • Notes

    Transferrins are iron binding transport proteins which can bind two Fe3+ ions in association with the binding of an anion, usually bicarbonate. It is responsible for the transport of iron from sites of absorption and heme degradation to those of storage and utilization. Serum transferrin may also have a further role in stimulating cell proliferation.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Transferrin Capture Antibody 1 x 600µl
    10X Human Transferrin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human Transferrin Lyophilized Purified Protein (ab91435) 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 2 x 50ml
    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
    • Cardiovascular
    • Blood
    • Other
    • Signal Transduction
    • Protein Trafficking
    • Organelle Proteins
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cardiovascular ELISA kits
  • Function

    Transferrins are iron binding transport proteins which can bind two Fe(3+) ions in association with the binding of an anion, usually bicarbonate. It is responsible for the transport of iron from sites of absorption and heme degradation to those of storage and utilization. Serum transferrin may also have a further role in stimulating cell proliferation.
  • Tissue specificity

    Expressed by the liver and secreted in plasma.
  • Involvement in disease

    Defects in TF are the cause of atransferrinemia (ATRAF) [MIM:209300]. Atransferrinemia is rare autosomal recessive disorder characterized by iron overload and hypochromic anemia.
  • Sequence similarities

    Belongs to the transferrin family.
    Contains 2 transferrin-like domains.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P02787 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Apotransferrin
    • Beta 1 metal binding globulin
    • Beta-1 metal-binding globulin
    • DKFZp781D0156
    • PRO1400
    • PRO1557
    • PRO2086
    • Serotransferrin
    • Serotransferrin precursor
    • Siderophilin
    • TF
    • TFQTL1
    • Transferin
    • Transferrin
    • TRFE_HUMAN
    see all
  • Database links

    • Entrez Gene: 7018 Human
    • Omim: 190000 Human
    • SwissProt: P02787 Human
    • Unigene: 518267 Human

    Associated products

    • ELISA kits

      • Human Transferrin ELISA Kit (ab108902)

    Images

    • Other - Human Transferrin ELISA Kit (ab187391)
      Other - Human Transferrin ELISA Kit (ab187391)

      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 Transferrin standard curve comparison
      Human Transferrin standard curve comparison

      Standard Curve comparison between the original Human Transferrin SimpleStep ELISA kit and current Human Transferrin SimpleStep ELISA kit.

    • Example of human Transferrin standard curve in Sample Diluent NS.
      Example of human Transferrin standard curve in Sample Diluent NS.

      The Transferrin 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.

    • Standard curve
      Standard curve

      Example of human Transferrin standard curve in Sample Diluent NS. The Transferrin standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native Transferrin in human serum and plasmas samples.
      Interpolated concentrations of native Transferrin in human serum and plasmas samples.

      The concentrations of Transferrin were measured in duplicates, interpolated from the Transferrin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:50,000, plasma (EDTA) 1:50,000, plasma (citrate) 1:50,000 and plasma (heparin) 1:50,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Transferrin concentration was determined to be 1.82 mg/mL in serum, 1.84 ng/mL in plasma (EDTA) and 1.86 mg/mL in plasma (citrate) and 1.43 ng/mL in plasma (heparin).

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

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Transferrin concentration was determined to be 2.38 mg/mL with a range of 1.54 – 3.2 mg/mL.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Native Transferrin was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.

    • Sandwich ELISA - Human Transferrin ELISA Kit (ab187391)
      Sandwich ELISA - Human Transferrin ELISA Kit (ab187391)
      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 (4)

    Publishing research using ab187391? Please let us know so that we can cite the reference in this datasheet.

    ab187391 has been referenced in 4 publications.

    • Harder NHO  et al. Effects of Dietary Glucose and Fructose on Copper, Iron, and Zinc Metabolism Parameters in Humans. Nutrients 12:N/A (2020). PubMed: 32854403
    • Vatine GD  et al. Human iPSC-Derived Blood-Brain Barrier Chips Enable Disease Modeling and Personalized Medicine Applications. Cell Stem Cell 24:995-1005.e6 (2019). PubMed: 31173718
    • Allden SJ  et al. The Transferrin Receptor CD71 Delineates Functionally Distinct Airway Macrophage Subsets during Idiopathic Pulmonary Fibrosis. Am J Respir Crit Care Med N/A:N/A (2019). PubMed: 31051082
    • Anantpadma M  et al. Large Scale Screening and Identification of Novel Ebolavirus and Marburgvirus Entry Inhibitors. Antimicrob Agents Chemother N/A:N/A (2016). Sandwich ELISA ; Human . PubMed: 27161622

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Question

    Do both the capture antibody and the detector antibody recognize hTF?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 05 2015

    Answer

    Yes, the capture and detector antibody recognize human Transferrin.

    Read More

    Karen Baker

    Abcam Scientific Support

    Answered on Mar 05 2015

    Question

    1) What is the immunogen of the antibody? Generated different stable cell lines with mutations in the transferrin protein (aa95, 188, 426, 517). Will these mutations affect antibody binding?

    2) Will the antibody/assay work equally well with transferrin bound to bicarbonate (naturally occuring) and oxycholate? See lower singal with transferrin bound to oxycholate.

    3) Is quantification affected by the amount of iron loaded onto the transferrin molecule?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 27 2015

    Answer

    1) The antibodies in this kit have been raised against native transferrin and unfortunately we do not know precise epitopes. A monoclonal capture antibody and a polyclonal detector antibody is provided. Therefore we do not know if the mutations generated will be recognized.

    2 and 3) We have not tested the effects of bicarbonate, oxycholate or iron on assay activity. The standard protein in the kit is native purified human transferrin. The kit works with this protein and with the human serum/plasma samples and has very good linearity of dilution. Therefore we expect that this antibody pair is compatible with normal levels of bicarbonate/oxycholate/iron bound to transferrin. However we do not know if abnormally high/low levels of any of these will affect the reactivity of the sample.

    Read More

    Karen Baker

    Abcam Scientific Support

    Answered on Feb 27 2015

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