Human Lactoferrin ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human Lactoferrin with a sensitivity of 17.7 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Images
Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 19.5 - 80000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 17.7 pg/mL
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
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Human Lactoferrin ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human Lactoferrin with a sensitivity of 17.7 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Urine, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 19.5 - 80000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 17.7 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 8 | C.V. 5.1 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample 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 Saliva | Average % = 120 | Range 113 - 126 % |
Sample type Urine | Average % = 99 | Range 99 - 100 % |
Sample type Heparin Plasma | Average % = 106 | Range 103 - 108 % |
Sample type Citrate plasma | Average % = 110 | Range 109 - 111 % |
Sample type Cell culture media | Average % = 86 | Range 86 - 86 % |
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
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.
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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4 product images
Sandwich ELISA - Human Lactoferrin ELISA Kit, Fluorescent (ab229392)
Example of human Lactoferrin standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Lactoferrin ELISA Kit, Fluorescent (ab229392)
Linearity of dilution of native Lactoferrin in serum and urine and spiked Lactoferrin in media.
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).
Sandwich ELISA - Human Lactoferrin ELISA Kit, Fluorescent (ab229392)
Linearity of dilution of native Lactoferrin in human plasmas.
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).
Sandwich ELISA - Human Lactoferrin ELISA Kit, Fluorescent (ab229392)
Levels of Lactoferrin in human donors.
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.
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