Overview

• Product nameHuman Lactoferrin ELISA Kit SimpleStep
  See all Lactoferrin kits
• Detection methodColorimetric
• Precision

  Intra-assay

  Sample	n	Mean	SD	CV%
  Human Serum	8			2.39%

• Sample type
  Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
• Assay typeSandwich (quantitative)
• Sensitivity
  123 pg/ml
• Range
  234 pg/ml - 15000 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%
  Cell culture media	114	113% - 116%
  Heparin Plasma	105	93% - 119%
  EDTA Plasma	117	115% - 120%
  Citrate Plasma	101	98% - 106%

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

  Abcam’s Lactoferrin Human in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Lactoferrin Human 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 applicationsSandwich ELISAmore details
• PlatformMicroplate (12 x 8 well strips)

Properties

• Storage instructionsStore 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	1 x 20ml
  Antibody Diluent CPI	1 x 6ml
  Human Lactoferrin Lyophilized Purified Protein	2 vials
  Plate Seals	1 unit
  Sample Diluent NS	1 x 50ml
  SimpleStep Pre-Coated 96-Well Microplate	1 unit
  Stop Solution	1 x 12ml
  TMB Substrate	1 x 12ml

• Research areas
  • Cardiovascular
  • Blood
  • Serum Proteins

  • Signal Transduction
  • Metabolism
  • Vitamins / Minerals

  • Cancer
  • Invasion/microenvironment
  • Apoptosis
  • Other

  • Cancer
  • Cell Death
  • Apoptosis
  • Other

Applications

Human Lactoferrin ELISA Kit SimpleStep images

• 

  Example of Human Lactoferrin standard curve.

  Background-subtracted data values (mean +/- SD) are graphed.

• 

  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).

• 

  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).

• 

  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.