Human Lactoferrin ELISA Kit

Sample

Human plasma, serum, milk, urine, saliva, CSF, cell culture, and cell lysate samples.

Species Reactivity

Human

Detection Method

sELISA

Intended Use

Human Lactoferrin ELISA (Enzyme-Linked Immunosorbent Assay) Kit is designed for detection of lactoferrin in human plasma, serum, milk, urine, saliva, CSF, cell culture, and cell lysate samples.

Contents of Kit

1.Human Lactoferrin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human lactoferrin.
2.Sealing Tapes: Each kit contains 3 precut, pressure sensitive sealing tapes that can be cut to fit the format of the individual assay.
3.Human Lactoferrin Standard: Human lactoferrin in a buffered protein base (42 ng, lyophilized).
4.Biotinylated Human Lactoferrin Antibody (50x): A 50-fold concentrated biotinylated polyclonal antibody against human lactoferrin (120 μl).
5.MIX Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml).
6.Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles).
7.SP Conjugate (100x): A 100-fold concentrate (80 μl).
8.Chromogen Substrate (1x): A stabilized peroxidase chromogen substrate tetramethylbenzidine (7 ml).
9.Stop Solution (1x): A 0.5 N hydrochloric acid solution to stop the chromogen substrate reaction (11 ml).

Storage

1.Upon arrival, immediately store components of the kit at recommended temperatures up to the expiration date.
2.Store SP Conjugate and Biotinylated Antibody at -20°C.
3.Store Microplate, Diluent Concentrate (10x), Wash Buffer, Stop Solution, and Chromogen Substrate at 2-8°C.
4.Unused microplate wells may be returned to the foil pouch with the desiccant packs and resealed. May be stored for up to 30 days in a vacuum desiccator.
5.Store Standard at 2-8°C before reconstituting with Diluent and at -20°C after reconstituting with Diluent.

Precision

The minimum detectable dose of human lactoferrin as calculated by 2SD from the mean of a zero standard was established to be 0.13 ng/ml.
• Intra-assay precision was determined by testing three plasma samples twenty times in one assay.
• Inter-assay precision was determined by testing three plasma samples in twenty assays.

General Description

Lactoferrin (lactotransferrin, growth-inhibiting protein 12, LTF) is an 80 kDa iron-binding glycoprotein produced by many exocrine glands with a major constituent in the secondary granules of neutrophilic leukocytes. It is found mainly in the oral cavity where it can come into direct contact with pathogens, such as viruses, bacteria, etc. Lactoferrin is present in maternal milk, saliva, tears, vaginal secretions, semen, bronchoalveolar lavage fluid, and specific granules of polymorphonuclear leukocytes [PMNs] (1). Serum lactoferrin concentration is much higher during inflammation (2). Lactoferrin is known to be an immune modulator or enhancer due to specific receptors for lactoferrin that are found on many key immune cells, such as lymphocytes, monocytes, and macrophages. It is also considered an antioxidant that scavenges free iron, helping to prevent uncontrolled iron-based free radical reactions, thus protecting certain cells from peroxidation. Lactoferrin is known to be directly involved in the up-regulation of natural killer (NK) cell activity (3). It directly inhibits viruses by binding to viral receptor sites, thus preventing the virus from infecting healthy cells. Lactoferrin has a direct bactericidal function to certain bacteria, such as Streptococcus mutans, Vibrio cholerae, Escherichia coli, Actinobacillus actinomycetemcomitans, and Legionella pneumophila; it also has a bacteriostatic effect that deprives iron-requiring bacteria of this essential growth nutrient (1, 3-4).

Standard Curve

The typical data is provided for reference only. Individual laboratory means may vary from the values listed. Variations between laboratories may be caused by technique differences.

The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.