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Catalog # HK357

Ficolin-1, Human, ELISA kit

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Ficolins are a family of pattern recognition molecules containing both a collagen-like domain and a fibrinogen-like domain that are capable of activating the complement system, through association with MASPs, on the surface of microorganisms. In human, three Ficolins are known: Ficolin-1 (M-ficolin), Ficolin-2 (L-ficolin), and Ficolin-3 (H-ficolin). Ficolin-1, is mainly expressed by monocytes, granulocytes and lung cells and is present locally in areas of inflammation. However, it was recently also found in serum suggesting that Ficolin-1 also has a role in systemic immunity like Ficolin-2 and Ficolin-3. Functional analysis of Ficolin-1 has demonstrated that it binds specifically to capsular polysaccharides of some microorganisms. Furthermore, it was shown that Ficolin-1 facilitates the uptake of E. coli by a leukemic monocyte lymphoma cell line, suggesting that it functions as a phagocytic receptor on the surface of circulating monocytes. Low cord blood concentration of Ficolin-1 has been associated with higher necrotizing enterocolitis-associated fatality in premature infants and polymorphisms in the Ficolin-1 gene are associated with the development of rheumatoid arthritis and susceptibility to leprosy.

Application
The human Ficolin-1 ELISA kit is to be used for the in vitro quantitative determination of human Ficolin-1 in serum and plasma samples.

Principle
The human Ficolin-1 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 a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are incubated in microtiter wells coated with antibodies recognizing human Ficolin-1. Biotinylated tracer antibody will bind to the captured human M-ficolin. 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 human Ficolin-1 standards (log). The human Ficolin-1 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Linearity
The linearity of the assay was determined by serially diluting plasma and serum samples containing human Ficolin-1. The diluted samples were measured in the assay.

Recovery
Normal human blood samples (plasma) containing baseline levels of human Ficolin-1, were spiked with human Ficolin-1, in concentrations of 2 and 0.2µg/ml. Samples with and without human Ficolin-1, were incubated for 1 hour at 37°C. Samples were measured using the ELISA. Recovery values for human Ficolin-1, ranged between 93.9% and 97.4%.

Product type
Assays
Quantity
1 x 96 det., 2 x 96 det.
Standard range
3.1 to 200 ng/ml
Detection level
3.1 ng/ml
Working volume
100 µl/well
Species
Human
Cross reactivity
Bovine – No, Canine – No, Goat – No, Horse – No, Mouse – No, Pig – No, Rabbit – No, Rat – No, Sheep – No
Alias
M-ficolin, ficolin/P35-related
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 of or derivation of this product.
Disease
Cardiology and metabolism, Infectious diseases
Safety Data Sheet Assay Manual
HK357
372 kb

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