Ficolin-2, Human, ELISA kit

Quantity:
1 x 96 det.
Catalog #:
HK336-01

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Ficolins are a group of proteins containing both a collagen-like domain and a fibrinogen-like domain. Three forms of ficolins have been identified in humans: Ficolin-1 (M-ficolin), Ficolin-2 (L-ficolin),and Ficolin-3 (H-ficolin). Ficolin-2 is produced primarily by the liver. Whereas Ficolin-3 is produced by the liver and the lung and Ficolin-1 by monocytes, leukocytes and lung cells. Ficolin-2 recognizes distinct danger-associated molecular patterns (DAMPs) like GlcNAc-structures in Lipoteichoic Acid (LTA) and fungal 1,3-β-D-glucan. Ficolin-2 also recognizes N-acetylated compounds such as acetylcholine. Furthermore, Ficolin-2 recognizes apoptotic cells and participates in the removal of host cells. Ficolin-2 circulates in complex with MASP-2 and can activate the lectin complement pathway. Ficolin-2 serum levels are reported with varying concentrations. Polymorphisms in the promoter of the FCN2 gene from which Ficolin-2 is derived are associated with variation in Ficolin-2 serum levels. These polymorphisms may influence the affinity of Ficolin-2 to GlcNAc-structures. Ficolin-2 deficiencies have not been reported, but low Ficolin-2 serum levels have been associated with recurrent respiratory infections in children. Interestingly, Ficolin-2 has been implicated in the unique immune challenge during pregnancy. In maternal plasma of normal pregnancies, a 4- to 5-fold increase in Ficolin-2 was detected compared to healthy non-pregnant persons. However, significantly lower Ficolin-2 maternal plasma concentrations were associated with pre-eclamptic pregnancies. Therefore, assessment of Ficolin-2 is warranted to study its regulatory role in the innate immune system. Normal human plasma contains a Ficolin-2 concentration ranging from 1 to 14 µg/ml.
Specifications
Product type Assays
Quantity 1 x 96 det.
Standard range 16-1000 ng/ml
Detection level 16 ng/ml
Working volume 100 µl/well
Species Human
Alias P35, Ficolin L, L-ficolin, EBP-37, hucolin, FCN2
Application The human Ficolin-2 ELISA kit is to be used for the in vitro quantitative determination of human Ficolin-2 in serum, plasma and cell culture supernatant samples.
Principle The human Ficolin-2 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are captured by a solid bound specific antibody. Biotinylated tracer antibody will bind to captured Ficolin-2. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the Ficolin-2 standards (log). The human Ficolin-2 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Storage and stability Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Precautions For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use or derivation of this product.
References 1. Atkinson, A et al; L-Ficolin in children with recurrent respiratory infections. Clin Exp Immunol 2004, 138: 517
2. Bidula, S et al; The serum opsonin L-ficolin is detected in lungs of human transplant recipients following fungal infections and modulates inflammation and killing of Aspergillus fumigatus. J Infect Dis 2015, 212: 234
3. Schlapbach, L et al; M-ficolin in the neonatal period: Associations with need for mechanical ventilation and mortality in premature infants with necrotising enterocolitis. Mol Immunol 2009, 46: 2597
Disease Cardiology and metabolism, Infectious diseases
Applications
Application assays: The human Ficolin-2 ELISA kit is to be used for the in vitro quantitative determination of human Ficolin-2 in serum, plasma and cell culture supernatant samples.
Principle: The human Ficolin-2 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are captured by a solid bound specific antibody. Biotinylated tracer antibody will bind to captured Ficolin-2. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the Ficolin-2 standards (log). The human Ficolin-2 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
References
Perfomance: The linearity of the assay was determined by serially diluting a sample containing 6903 ng/ml human Ficolin-2. The diluted samples were measured in the assay. The line obtained a slope of 1.11 and a correlation coefficient of 0.998.
Recovery: Normal human blood samples (plasma), containing baseline levels of 500 ng/ml were spiked with recombinant Ficolin-2 in concentrations of 20 and 200 ng/ml. Samples with and without Ficolin-2 were incubated for 1 hour at 37 °C. Samples were measured using the ELISA. Recovery values for Ficolin-2 ranged between 100 % and 110%.
References
References: 1. Atkinson, A et al; L-Ficolin in children with recurrent respiratory infections. Clin Exp Immunol 2004, 138: 517
2. Bidula, S et al; The serum opsonin L-ficolin is detected in lungs of human transplant recipients following fungal infections and modulates inflammation and killing of Aspergillus fumigatus. J Infect Dis 2015, 212: 234
3. Schlapbach, L et al; M-ficolin in the neonatal period: Associations with need for mechanical ventilation and mortality in premature infants with necrotising enterocolitis. Mol Immunol 2009, 46: 2597
Assay Manual
pdf
(Size: 0.61 MB)
Safety Data Sheet
pdf
(Size: 0.14 MB)
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