Overview

  • Product name
    Human Lactoferrin ELISA Kit, Fluorescent
    See all Lactoferrin kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 8 2.39%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 10.68%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    17.2 pg/ml
  • Range
    23 pg/ml - 12000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 110 109% - 111%
    Milk 109 96% - 116%
    Urine 102 89% - 112%
    Serum 102 96% - 106%
    Plasma 105 93% - 119%
    Cell culture media 114 113% - 116%
    EDTA Plasma 117 115% - 120%
    Citrate Plasma 101 98% - 106%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Rabbit, Goat, Guinea pig, Hamster, Cow
  • Product overview

    Lactoferrin in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Lactoferrin protein in human serum, plasma, milk, urine, saliva, and cell culture supernatants.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • 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
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Human Lactoferrin Capture Antibody 1 x 600µl
    10X Human Lactoferrin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent CPI 1 x 6ml
    Human Lactoferrin Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 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 ab229392 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
  • Background-subtracted data values (mean +/- SD) are graphed.

  • Native human Lactoferrin protein was measured in serum diluted 1:64 and urine diluted 1:8 in Sample Diluent NS prior to a 2-fold dilution series in Sample Diluent NS. Purified human Lactoferrin protein was spiked into cell culture media diluted 1:20 prior to a fold dilution series in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Native human Lactoferrin protein was measured in citrate plasma (1:128), EDTA plasma (1:150), and heparin plasma (1:64) diluted in a 2-fold dilution series in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Ten individual healthy male donors and one pooled sample (n = 50) were evaluated for the presence of Lactoferrin in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected by sample dilution (1:75). In the 10 individual donors, the mean level of Lactoferrin is 350.3 ng/mL with a range of 106.2 – 627.7 ng/mL and a standard deviation of 159.4 ng/mL.

Protocols

References

ab229392 has not yet been referenced specifically in any publications.

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