Book a Meeting

Factor H, 402H/Y Variant ELISA Kit [Human], 1 x 96 det. (CAT#: CB-P109-K) Datasheet

Product Type
Kit
Description
Complement is the main defense system of innate immunity, designed to eliminate microorganisms. There are three ways to activate complement. The classical pathway is initiated by immune complexes, the lectin pathway is initiated by surface-bound mannan-bound lectins, and the alternative pathway is initiated by all surfaces that are not protected. One of the central complement regulators of the alternative pathway is complement factor H (CFH). CFH is the main soluble inhibitor of the alternative pathway of the complement system and plays a key role in controlling complement activation in the body. CFH is a 150 kDa plasma protein, mainly produced in the liver, and is present in the plasma at a concentration of about 500 μg/ml. The molecule consists entirely of a string of 20 folded globular domains called short consensus repeats (SCR). The activation of the alternative pathway is caused by the failure of spontaneous renewal to properly regulate the constant low levels of the abundant complement C3 to C3 (H2O). The surface of non-complement activation cells and other hosts are protected by CFH binding from other complement attacks. CFH inhibits the bypass by binding to C3b and reducing the formation and activation of the bypass C3 convertase (C3bBb). It also accelerates the degradation of this invertase and acts as a cofactor for serine protease factor I, degrading C3b into inactive C3b. CFH is associated with several diseases, such as atypical hemolytic syndrome (aHUS), membranous proliferative glomerulonephritis type II (MNGN) and age-related macular degeneration (AMD). In the Western world, AMD is the main cause of natural and irreversible blindness among the elderly, affecting 50 million people worldwide. It has been determined that the common polymorphism of the CFH gene on chromosome 1 is closely related to the risk of human suffering from 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 the CFH region that binds heparin and C-reactive protein. H402 (genotype CC; CFH-HH402) homozygous individuals have a 12-fold increased risk of AMD, while heterozygotes (CT genotype; CFH-HY402) are compared with 402Y homozygous individuals (AMD genotype; CFH-HY402). The risk of AMD is 2.5 times greater. Genotype TT; CFH-YY402). The CFH-HH402 and CFH-HY402 variants may explain approximately 43% of AMD in the elderly.
Applications
Complement factor H, H402 and Y402 variant detection ELISA kits are used for in vitro detection of H402 and Y402 variants of complement factor H in serum and plasma samples.
Principle
The ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 1 hour. The effective format of 1 plate has 3 (CFH, CFH-H402 and CFH-Y402) × 4 disposable 8-well plates, and the batch size can be freely selected for determination. Incubate the sample and control with a peroxidase-conjugated secondary antibody (conjugate) in a microtiter well coated with CFH (colorless), CFH H402 (white) and CFH Y402 (Black) Antibodies for all variants. During the incubation, CFH, CFH H402 or CFH Y402 are captured by the consolidated anti-cranial antibody. The bound antibody will bind to the captured CFH variant. The peroxidase-conjugated antibody will react with the substrate tetramethylbenzidine (TMB). The enzyme reaction is stopped by adding oxalic acid. Measure the absorbance at 450nm with a spectrophotometer. The polymorphism of complement factor H (Y402H) can be determined from the control.
Size
1 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
Detection of H402 and Y402 variants of complement factor H.
Sample
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Specimen Collection
Blood samples were collected using sterile venipuncture techniques, and serum was obtained using standard procedures. At least 5 ml of whole blood is recommended. Let the blood clot in the serum test tube for 60-65 minutes at room temperature (20-25°C). Centrifuge the blood sample and transfer the cell-free serum to a clean test tube. The serum must be handled correctly to prevent complement activation in vitro. Serum should be frozen in a sealed tube at -70°C or lower for long-term storage or transport on dry ice. The sample should not be frozen and thawed more than once. Do not use jaundice, lipemia and hemolytic serum. You cannot use heat-inactivated serum. Plasma cannot be used.
Sample Volume
50 µl/well
Species Reactivity
Human
Protocol Length
2 hours
Storage
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Research Use
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.

All products and services are for Research Use Only. Do Not use in humans.

ONLINE INQUIRY

Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.

  • *
  • *
  • *
USA

UK