Human Lactoferrin ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Lactoferrin with a sensitivity of 48.8 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
- Cited in over 15 citations
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- PloS one 18:e02912972023Salivary concentrations of secretory leukocyte protease inhibitor and matrix metallopeptidase-9 following a single bout of exercise are associated with intensity and hydration status.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKaren Knipping,Shirley W Kartaram,Marc Teunis,Nicolaas P A Zuithoff,Nicole Buurman,Laura M'Rabet,Klaske van Norren,Renger Witkamp,Raymond Pieters,Johan GarssenPubMed 37992002
- Scientific reports 13:49032023The abundances of LTF and SOD2 in amniotic fluid are potential biomarkers of gestational age and preterm birth.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTe-Yao Hsu,Hsin-Hsin Cheng,Kuo-Chung Lan,Hsuan-Ning Hung,Yun-Ju Lai,Chih-Chang Tsai,Wen-Lang Fan,Sung-Chou LiPubMed 36966172
- The Tohoku journal of experimental medicine 259:135-1422022Exploratory Study of Serum Lactoferrin and Anti-Lactoferrin Antibody Concentrations in Patients with Endometriosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKaya Mori-Yamanaka,Fuminori Kimura,Akiko Nakamura,Tetsuro Hanada,Jun Kitazawa,Aina Morimune,Shunichiro Tsuji,Takashi MurakamiPubMed 36476585
- International journal of molecular sciences 23:2022Possible Implications of Bacteriospermia on the Sperm Quality, Oxidative Characteristics, and Seminal Cytokine Network in Normozoospermic Men.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEva Tvrdá,Daniel Lovíšek,Eliška Gálová,Marianna Schwarzová,Eva Kováčiková,Simona Kunová,Jana Žiarovská,Miroslava KačániováPubMed 35955814
- Investigative ophthalmology & visual science 63:102022Targeted Analysis of Tears Revealed Specific Altered Metal Homeostasis in Age-Related Macular Degeneration.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEva Valencia,Montserrat García,Beatriz Fernández-Vega,Rosario Pereiro,Lara Lobo,Héctor González-IglesiasPubMed 35426907
- Alzheimer's research & therapy 13:1502021Salivary lactoferrin is associated with cortical amyloid-beta load, cortical integrity, and memory in aging.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLucia Reseco,Mercedes Atienza,Marina Fernandez-Alvarez,Eva Carro,Jose L CanteroPubMed 34488875
- BMC oral health 21:3852021Origin of MMP-8 and Lactoferrin levels from gingival crevicular fluid, salivary glands and whole saliva.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLiza L Ramenzoni,Deborah Hofer,Alex Solderer,Daniel Wiedemeier,Thomas Attin,Patrick R SchmidlinPubMed 34353321
- Alzheimer's & dementia (Amsterdam, Netherlands) 13:e121732021Standardizing salivary lactoferrin measurements to obtain a robust diagnostic biomarker for Alzheimer's disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesFernando Bartolome,Gorka Orive,Eva CarroPubMed 33969170
- Cell 182:1044-1061.e182020Extracellular Vesicle and Particle Biomarkers Define Multiple Human Cancers.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAyuko Hoshino et. al.PubMed 32795414
- EBioMedicine 57:1028342020Decreased salivary lactoferrin levels are specific to Alzheimer's disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMarta González-Sánchez,Fernando Bartolome,Desiree Antequera,Veronica Puertas-Martín,Pilar González,Adolfo Gómez-Grande,Sara Llamas-Velasco,Alejandro Herrero-San Martín,David Pérez-Martínez,Alberto Villarejo-Galende,Mercedes Atienza,Miriam Palomar-Bonet,Jose Luis Cantero,George Perry,Gorka Orive,Borja Ibañez,Hector Bueno,Valentin Fuster,Eva CarroPubMed 32586758
Key facts
- Detection method
- Colorimetric
- Sample types
- Saliva, Urine, Cell culture media, Heparin Plasma, Citrate plasma, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 156.3 - 10000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 48.8 pg/mL
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Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
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. Major iron-binding and multifunctional protein found in exocrine fluids such as breast milk and mucosal secretions (PubMed:11179314, PubMed:12693969, PubMed:14573629, PubMed:1599934, PubMed:3169987, PubMed:6802759). Has antimicrobial activity, which depends on the extracellular cation concentration (PubMed:6802759). Antimicrobial properties include bacteriostasis, which is related to its ability to sequester free iron and thus inhibit microbial growth, as well as direct bactericidal properties leading to the release of lipopolysaccharides from the bacterial outer membrane (PubMed:11179314, PubMed:12693969, PubMed:14573629, PubMed:1599934, PubMed:3169987, PubMed:6802759). Can also prevent bacterial biofilm development in P.aeruginosa infection (PubMed:12037568). Has weak antifungal activity against C.albicans (PubMed:11083624). Has anabolic, differentiating and anti-apoptotic effects on osteoblasts and can also inhibit osteoclastogenesis, possibly playing a role in the regulation of bone growth (PubMed:15166119). Promotes binding of species C adenoviruses to epithelial cells, promoting adenovirus infection (PubMed:17079302). Can inhibit papillomavirus infections (PubMed:17481742). Stimulates the TLR4 signaling pathway leading to NF-kappa-B activation and subsequent pro-inflammatory cytokine production while also interfering with the lipopolysaccharide (LPS)-stimulated TLR4 signaling (PubMed:20345905). Inhibits neutrophil granulocyte migration to sites of apoptosis, when secreted by apoptotic cells (PubMed:19033648). Stimulates VEGFA-mediated endothelial cell migration and proliferation (PubMed:16842782). Binds heparin, chondroitin sulfate and possibly other glycosaminoglycans (GAGs) (PubMed:9359845). Also binds specifically to pneumococcal surface protein A (PspA), the lipid A portion of bacterial lipopolysaccharide (LPS), lysozyme and DNA (PubMed:9359845). Lactoferricin binds to the bacterial surface and is crucial for the bactericidal functions. Has some antiviral activity against papillomavirus infection (PubMed:17481742). N-terminal region shows strong antifungal activity against C.albicans (PubMed:11083624). Contains two BBXB heparin-binding consensus sequences that appear to form the predominate functional GAG-binding site. Kaliocin-1. Has antimicrobial activity and is able to permeabilize different ions through liposomal membranes. Lactoferroxin-A. Has opioid antagonist activity (PubMed:1369293). Shows preference for mu-receptor (PubMed:1369293). Lactoferroxin-B. Has opioid antagonist activity (PubMed:1369293). Shows higher degrees of preference for kappa-receptors than for mu-receptors (PubMed:1369293). Lactoferroxin-C. Has opioid antagonist activity (PubMed:1369293). Shows higher degrees of preference for kappa-receptors than for mu-receptors (PubMed:1369293). The lactotransferrin transferrin-like domain 1 functions as a serine protease of the peptidase S60 family that cuts arginine rich regions (PubMed:12535064). This function contributes to the antimicrobial activity (PubMed:12535064). Shows a preferential cleavage at -Arg-Ser-Arg-Arg-|- and -Arg-Arg-Ser-Arg-|-, and of Z-Phe-Arg-|-aminomethylcoumarin sites (PubMed:12535064). Isoform DeltaLf. Transcription factor with antiproliferative properties and ability to induce cell cycle arrest (PubMed:15222485). Binds to the DeltaLf response element found in the SKP1, BAX, DCPS, and SELENOH promoters (PubMed:22320386).
Alternative names
GIG12, LF, LTF, Lactotransferrin, Lactoferrin, Growth-inhibiting protein 12, Talalactoferrin
What's included?
Recommended products
Human Lactoferrin ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Lactoferrin with a sensitivity of 48.8 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
- Cited in over 15 citations
Key facts
- Detection method
- Colorimetric
- Sample types
- Saliva, Urine, Cell culture media, Heparin Plasma, Citrate plasma, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 156.3 - 10000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate (12 x 8 well strips)
- Sensitivity
- = 48.8 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 8 | C.V. 5.1 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 3 | C.V. 5.4 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Milk | Average % = 109 | Range 99 - 118 % |
Sample type Serum | Average % = 102 | Range 100 - 105 % |
Sample type EDTA Plasma | Average % = 108 | Range 105 - 110 % |
Sample type Urine | Average % = 99 | Range 99 - 100 % |
Sample type Saliva | Average % = 120 | Range 113 - 126 % |
Sample type Cell culture media | Average % = 86 | Range 86 - 86 % |
Sample type Heparin Plasma | Average % = 106 | Range 105 - 110 % |
Sample type Citrate plasma | Average % = 110 | Range 109 - 111 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
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 (Pre-coated 384 well Microplate SimpleStep ELISA® 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
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).
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Lactoferrin also known as lactotransferrin is an iron-binding glycoprotein with a molecular weight of about 80 kDa. It predominantly exists in mammalian milk particularly in high concentrations in colostrum and is also found in various epithelial secretions such as saliva tears and nasal fluids. The liver and neutrophils also produce lactoferrin and it plays a significant role in the body's primary defense system. As an antibacterial agent lactoferrin sequesters free iron to limit bacterial growth and disrupt biofilms assisting in the protection against microbial invasions.
- Biological function summary
Lactoferrin exhibits multiple functions beyond its antimicrobial properties. It contributes to immunomodulation by binding to specific receptors on immune cells enhancing phagocytosis and influencing cytokine production. Also it possesses anti-inflammatory and antioxidant properties. Lactoferrin interacts with cellular components including glycosaminoglycans and lipoproteins impacting the activity and expression of proteins within signaling and metabolic pathways. This multifaceted role places it as a critical participant in both innate and adaptive immune responses although it does not form part of a stable protein complex.
- Pathways
Lactoferrin plays a significant role in the iron metabolism and immune response pathways. It tightly interacts with the transferrin receptor modulating iron uptake and distribution across tissues which helps prevent iron overload and oxidative stress. Furthermore lactoferrin links with the toll-like receptor pathways particularly TLR4 which enables the modulation of innate immune responses against pathogens. By influencing nuclear factor kappa B (NF-kB) signaling lactoferrin impacts inflammation and immune responses working together with proteins such as ferritin and hepcidin.
- Associated diseases and disorders
Lactoferrin has relevancy to diseases like anemia of chronic disease and inflammatory bowel disease (IBD). Its role in regulating iron homeostasis makes it essential for preventing anemia associated with chronic inflammation. Lactoferrin mitigates dysregulated immune responses in IBD potentially reducing inflammation and promoting gut health. Disease-modifying properties of lactoferrin connect it to glycoproteins like transferrin and inflammatory mediators demonstrating its potential therapeutic importance.
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Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
Example of human Lactoferrin standard curve in Sample Diluent NS.
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.
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
Interpolated concentrations of native Lactoferrin in human serum and plasma samples.
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).
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
Interpolated concentrations of native Lactoferrin in human milk (defatted), urine, and saliva samples.
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.
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
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 Lactoferrin concentration was determined to be 473.8 ng/mL with a range of 315.2 – 852.7 ng/mL.
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
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 Lactoferrin was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
Example of human Lactoferrin standard curve in Sample Diluent NS. The Lactoferrin standard curve was prepared. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Lactoferrin ELISA Kit (ab200015)
Linearity of dilution.
Native Lactoferrin was measured in human milk, urine, and saliva samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Native purified Lactoferrin protein was spiked into RPMI 1640 cell culture media containing 10% fetal bovine serum and measured in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
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