Overview

  • Product name

    Human Lactoferrin ELISA Kit
    See all Lactoferrin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human Serum 8 5.1%
    Inter-assay
    Sample n Mean SD CV%
    Human Serum 3 5.4%
  • Sample type

    Saliva, Milk, Urine, Serum, Cell culture media, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    48.8 pg/ml
  • Range

    156.3 pg/ml - 10000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 120 113% - 126%
    Milk 109 99% - 118%
    Urine 99 99% - 100%
    Serum 102 100% - 105%
    Cell culture media 86 86% - 86%
    Heparin Plasma 106 105% - 110%
    EDTA Plasma 108 105% - 110%
    Citrate Plasma 110 109% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow, Monkey
  • Product overview

    As of October 11 2019, Human Lactoferrin SimpleStep ELISA® kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency when quantifying Lactoferrin protein in serum, plasma, milk, urine, saliva and cell culture supernatant samples.


    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.

  • Notes

    Transferrins are iron binding transport proteins which can bind two Fe3+ ions in association with the binding of an anion, usually bicarbonate. Lactoferrin is a member of the transferrin family and is a major iron-binding and multifunctional protein found in exocrine fluids such as breast milk and mucosal secretions. High levels of Lactoferrin are found in saliva and tears, intermediate levels in serum and plasma, and low levels in urine. The canonical sequence of Lactoferrin (known as isoform 1) is 710 amino acids in length and runs at 78.2 kDa. Isoform Delta-Lactoferrin (or DeltaLF) is missing amino acids 1 – 44 of the canonical sequence. Mouse and rat Lactoferrin are 71% and 62% identical to human Lactoferrin, respectively.

    Lactoferrin possesses antibacterial, antiviral, anti-inflammatory, and immunoregulatory properties. The presence of Lactoferrin in neutrophils and the release of Lactoferrin during inflammation suggest that Lactoferrin is involved in phagocytic killing and immune responses. Additionally, Lactoferrin mediates bacteriostasis through its ability to sequester free iron and inhibit microbial growth. Lactoferrin also directs bactericidal properties leading to the release of lipopolysaccharides from the bacterial outer membrane. Furthermore, Lactoferrin may play a role in the regulation of bone growth because it has anabolic, differentiating, and anti-apoptotic effects on osteoblasts and can also inhibit osteoclastogenesis. Salivary Lactoferrin plays an important role in protecting the oral cavity against several oral pathogens including Streptococcus mutans. Additionally, increased levels of Lactoferrin in fecal samples is used as a marker of active inflammatory bowel disease (IBD).

  • 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 Lactoferrin Capture Antibody 1 x 600µl
    10X Human Lactoferrin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human Lactoferrin Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    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
  • Research areas

  • Function

    Transferrins are iron binding transport proteins which can bind two Fe(3+) ions in association with the binding of an anion, usually bicarbonate.
    Lactotransferrin has antimicrobial activity which depends on the extracellular cation concentration.
    Lactoferroxins A, B and C have opioid antagonist activity. Lactoferroxin A shows preference for mu-receptors, while lactoferroxin B and C have somewhat higher degrees of preference for kappa-receptors than for mu-receptors.
    The lactotransferrin transferrin-like domain 1 functions as a serine protease of the peptidase S60 family that cuts arginine rich regions. This function contributes to the antimicrobial activity.
  • Sequence similarities

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

    Secreted.
  • Information by UniProt
  • Alternative names

    • GIG12
    • Growth inhibiting protein 12
    • HLF2
    • Kaliocin 1
    • Lactoferrin
    • Lactoferroxin-C
    • Lactotransferrin
    • LF
    • LTF
    • Neutrophil lactoferrin
    • Talalactoferrin
    • TRFL_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab200015 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

  • 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.

     

  • The Lactoferrin 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 Lactoferrin were measured in duplicates, interpolated from the Lactoferrin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:200, plasma (citrate) 1:200, plasma (heparin) 1:200, and plasma (EDTA) 1:200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lactoferrin concentration was determined to be 1,142 ng/mL in neat serum, 885.1 ng/mL in neat plasma (citrate), 1,054 ng/mL in neat plasma (heparin), and 581.4 ng/mL in neat plasma (EDTA).  

  • The concentrations of Lactoferrin were measured in duplicates, interpolated from the Lactoferrin standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 1:500,000, urine 1:20, and saliva 1:10,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lactoferrin concentration was determined to be 4,021.6 µg/mL in neat milk (defatted), 0.033 µg/mL in neat urine, and 39.1 µg/mL in neat saliva.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lactoferrin concentration was determined to be 473.8 ng/mL with a range of 315.2 – 852.7 ng/mL.

Protocols

References

This product has been referenced in:

See 1 Publication for this product

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