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Human Lactoferrin ELISA Kit

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Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 48.8 pg/ml
  • Range: 156.3 pg/ml – 10000 pg/ml
  • Sample type: Cell culture media, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

Overview

  • Product name

    Human Lactoferrin ELISA Kit

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    SamplenMeanSDCV%
    Human Serum8  5.1%
    Inter-assay
    SamplenMeanSDCV%
    Human Serum3  5.4%

  • Sample type

    Saliva, Milk, Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    48.8 pg/ml

  • Range

    156.3 pg/ml – 10000 pg/ml

  • Recovery

     

    Sample specific recovery
    Sample typeAverage %Range
    Saliva120113% – 126%
    Milk10999% – 118%
    Urine9999% – 100%
    Serum102100% – 105%
    Cell culture media8686% – 86%
    Hep Plasma106105% – 110%
    EDTA Plasma108105% – 110%
    Cit plasma110109% – 111%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human
    Does not react with: Cow

  • Product overview

    As of October 11 2019, Human Lactoferrin ELISA kit has been re-developed. We have identified new recombinant monoclonal antibodies to provide improved performance and consistency.


    Human Lactoferrin ELISA Kit (ab200015) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Lactoferrin protein in edta plasma, hep plasma, milk, saliva, serum, urine, cell culture media, and cit plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Lactoferrin with 48.8 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.


    ASSAY SPECIFICITY


    This kit recognizes native human Lactoferrin protein in serum, plasma, milk, urine, saliva, and cell culture supernatant samples only.


    Cell and tissue extract samples have not been tested with this kit.


    CROSS REACTIVITY


    Recombinant human Transferrin protein was prepared at within the working range of the Lactoferrin standard curve and assayed for cross reactivity. No cross-reactivity was observed.


    INTERFERENCE


    Recombinant human Transferrin protein was prepared at 50 ng/mL and 2 ng/mL and assayed for interference in the presence 2 ng/mL human Lactoferrin. No Interference was observed. 


    SPECIES REACTIVITY


    This kit recognizes human Lactoferrin protein.


    Other species reactivity was determined by measuring 0.5% 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, Monkey


     

  • 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).

  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components1 x 96 tests
    10X Human Lactoferrin Capture Antibody1 x 600µl
    10X Human Lactoferrin Detector Antibody1 x 600µl
    10X Wash Buffer PT (ab206977)1 x 20ml
    Antibody Diluent 5BI1 x 6ml
    Human Lactoferrin Lyophilized Purified Protein2 vials
    Plate Seals1 unit
    Sample Diluent NS (ab193972)1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978)1 unit
    Stop Solution1 x 12ml
    TMB Development Solution1 x 12ml

  • Research areas

       
    • Blood
      •  
    • Serum Proteins
    • Signal Transduction
      •  
    • Metabolism
      •  
    • Vitamins / Minerals
    • Cancer
      •  
    • Invasion/microenvironment
      •  
    • Apoptosis
      •  
    • Other
    • Cancer
      •  
    • Cell Death
      •  
    • Apoptosis
      •  
    • Other

  • 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.

Protocols

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