Factor H, 402H/Y variant detection, Human, ELISA kit

Quantity:
1 x 96 det.
Catalog #:
HK353

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Complement is a major defense system of innate immunity and aimed to destroy microbes. There are three pathways of complement activation. The classical pathway is initiated by immune complexes, the lectin pathway by surface bound mannan binding lectin, and the alternative pathway by all the surfaces that are not protected against it. One of the central complement regulators of the alternative pathway is complement factor H (CFH). CFH is the major soluble inhibitor of the alternative pathway of the complement system and plays a key role in controlling complement activation in vivo. CFH is a 150-kDa plasma protein that is mainly produced in the liver and is present in plasma at a concentration of approximately 500 µg/ml. The molecule is made up entirely of a string of 20 folded globular domains known as short consensus repeats (SCRs). Alternative pathway activation results from a failure to appropriately regulate the constant low level of the abundant complement complement C3 to C3(H2O) due to spontaneous turnover. Complement non-activating cells and other host surfaces are protected from alternative pathway complement attack through binding of CFH. CFH inhibits the alternative pathway by binding C3b and reducing the formation and activation of the alternative pathway C3-convertase (C3bBb). It also accelerates the decay of this convertase and works as a cofactor for the serine proteinase factor I in the degradation of C3b into inactive C3b. CFH is associated with several diseases like atypical hemolytic syndrome (aHUS), membranoproliferative glomerulonephritis type ll (MNGN) and age-related macular degeneration (AMD). In the western world, AMD is the leading cause of natural irreversible blindness in the elderly, affecting 50 million individuals worldwide. A common polymorphism in the CFH gene on chromosome 1, has been determined to be strongly associated with a person's risk for developing AMD. The CFH gene polymorphism is characterized by a T-to-C single nucleotide substitution resulting in a tyrosine-histidine change at amino acid position 402 . The sequence change is in a region of CFH that binds heparin and C-reactive protein. Individuals homozygous for H402 (genotype CC; CFH-HH402) have up to 12-fold increased risk for developing AMD, while heterozygotes (genotype CT; CFH-HY402) have a 2.5-fold greater risk for developing AMD than individuals homozygous for 402Y (genotype TT; CFH-YY402). The CFH-HH402 and CFH-HY402 variants, likely explain about 43% of AMD in older adults.
Specifications
Product type Assays
Quantity 1 x 96 det.
Standard range Detection of H402 and Y402 variants of complement factor H.
Working volume 50 µl/well
Species Human
Alias CFHL1; CFHL3; HF1; HF2; HUS; MGC88246; beta-1-H-globulin
Application The complement factor H, H402 and Y402 variant detection ELISA kit is to be used for the in vitro detection of H402 and Y402 variants of complement factor H in serum and plasma samples.
Principle The HK353 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 1½ hours. The efficient format of 1 plate with 3 (CFH, CFH-H402 and CFH-Y402) x 4 disposable 8-well strips allows free choice of batch size for the assay. Samples and controls are incubated together with peroxidase-conju­gated secondond antibody (conjugate) in microtiter wells coated with antibodies recognizing all variants of CFH (colorless), CFH H402 (white) and CFH Y402 (black). During incubation CFH, CFH H402 or CFH Y402 are captured by the solid bound anti­body. The conjugated antibodies will bind to the captured CFH variants. The peroxidase conjugated antibody 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. The polymorphism (Y402H) of complement factor H can be determined from the controls.
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 from the use or derivation of this product.
References 1. Hakobyan, S et al; Measurement of factor H variants in plasma using variant-specific monoclonal antibodies: application to accessing risk of age-related macular degeneration. Invest Ophthalmol Vis Sci 2008, 49: 1983
2. Szarvas, N et al; First-line therapy in atypical hemolytic uremic syndrome: consideration on infants with a poor prognosis. Ital J Pediatrics 2014, 40:101
Disease Autoimmunity, Nephrology
Applications
Application assays: The complement factor H, H402 and Y402 variant detection ELISA kit is to be used for the in vitro detection of H402 and Y402 variants of complement factor H in serum and plasma samples.
Principle: The HK353 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 1½ hours. The efficient format of 1 plate with 3 (CFH, CFH-H402 and CFH-Y402) x 4 disposable 8-well strips allows free choice of batch size for the assay. Samples and controls are incubated together with peroxidase-conju­gated secondond antibody (conjugate) in microtiter wells coated with antibodies recognizing all variants of CFH (colorless), CFH H402 (white) and CFH Y402 (black). During incubation CFH, CFH H402 or CFH Y402 are captured by the solid bound anti­body. The conjugated antibodies will bind to the captured CFH variants. The peroxidase conjugated antibody 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. The polymorphism (Y402H) of complement factor H can be determined from the controls.
References
References: 1. Hakobyan, S et al; Measurement of factor H variants in plasma using variant-specific monoclonal antibodies: application to accessing risk of age-related macular degeneration. Invest Ophthalmol Vis Sci 2008, 49: 1983
2. Szarvas, N et al; First-line therapy in atypical hemolytic uremic syndrome: consideration on infants with a poor prognosis. Ital J Pediatrics 2014, 40:101
Assay Manual
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(Size: 0.52 MB)
Safety Data Sheet
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