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Catalog # HK355

Complement factor I, Human, ELISA kit

The human Complement FI ELISA kit is to be used for the in vitro quantitative determination of FI in serum and plasma samples. Recently newly developed and improved, making the assay faster and more robust.

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825.001,359.00

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Complement Factor I (FI) is a serine protease, essential for the regulation of the complement system, and is encoded by the FI gene on chromosome 4. Known as the C3b/C4b inactivator, FI plays a crucial role in modulating complement activation by cleaving C3b and C4b in both cell-bound and fluid phases. Predominantly synthesized in the liver, FI begins as an 88 kDa
precursor protein that is processed into its active form—a disulfide-linked dimer composed of a 51 kDa heavy chain and a 37 kDa light chain.
Genetic diversity in the FI gene, including polymorphisms like R201S, R406H, and R502L, contributes to variations in FI function. A deficiency in FI can lead to decreased plasma levels of complement component 3 (C3), due to unchecked activation of the complement alternative pathway. This can result in recurrent bacterial infections, particularly in children. Additionally, recent findings have implicated mutations in the FI gene in the pathogenesis of Haemolytic Uremic Syndrome, a serious renal condition also linked to uncontrolled complement activity.

Application
The complement factor I ELISA kit is to be used for the in vitro quantitative determination of human CFI in plasma and serum.

Principle
The human FI ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 1 hour and 15 minutes. 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 FI. Peroxidase-conjugated antibody will bind to the captured FI. 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 FI standards (log). The human FI concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Recovery
Recovery was determined by mixing FI depleted serum with different EDTA plasma samples & using four EDTA plasma samples with a known FI concentration. Samples were incubated for one hour at room temperature and measured using the ELISA. The recovery experiment met the requirements and ranged between 87% and 108%

Precision and reproducibility
The intra-assay precision and reproducibility was tested using four samples with three different dilutions. CV % ranged between 1.6 and 5.5%. To determine the inter-assay variation, four samples were tested by two operators both on three separate days. CV % ranged between 3.6 and 5.8%.

Product type
Assays
Amount
1 x 96 det., 2 x 96 det.
Standard range
1.56 to 100 ng/ml
Detection level
Minimum concentration which can be measured is 1.56 ng/ml.
Working volume
100 µl/well
Species
Human
Cross reactivity
Mouse – No, Pig – No, Rat – No
Alias
CFI, C3b-Inactivator, C3b/C4b inactivator, Konglutinogen-Activating Factor
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.
Safety Data Sheet Assay Manual
HK355
222 kb

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