Complement factor H ELISA Kit [Human], 1 x 96 det. (CAT#: CB-P111-K) Datasheet

Product Type
Kit
Description
Complement factor H (CFH) is the first regulatory protein of the alternative pathway of the complement system. There are three ways to activate complement. The classical pathway is triggered by immune complexes. Surface-bound mannan-bound lectin pathways; and all surfaces that are not specifically protected. Each produces C3 convertase, a serine protease that cleaves the central complement protein C3 and produces the main cleavage fragment C3b. The complement system mediates many basic biological functions and participates in the host's resistance to infection, the initiation of inflammatory responses, the processing and clearance of immune complexes, and the regulation of immune responses. CFH is a 150 kD single-chain serum glycoprotein with a modular structure consisting of 20 homologous units of about 60 amino acids in series, called short consensus repeat (SCR). Many functional sites have been identified along the 20 SCR domain structure of Factor H. In SCR6-10 and SCR13-20 in SCR1-4. Three polyanionic binding sites have also been identified in SCR7, 13 and 20, such as heparin and several glycosaminoglycans. FH shows anti-inflammatory function and acts as a ligand for CRP. CFH has two important functional domains located at opposite ends of the protein. The N-terminal fragment of the factor H molecule is an essential liquid phase regulator for the alternative pathway. CFH binds to the surface of cells and tissues through the C-terminal domain and SCR 7, thereby mediating its protective effect on the surface of host cells. CFH is a relatively abundant plasma protein with a concentration of 400-800μg/ml, which is essential for maintaining the homeostasis of complement and limiting the effect of complement on activated surfaces. CFH binds to C3b, accelerates the decay of bypass C3 convertase (C3bBb), and acts as a cofactor for factor I-mediated C3b proteolytic inactivation. Factor H regulates complement on the liquid phase and on the cell surface. Genetic analysis shows that there is a clear association between CFH and different human diseases. These diseases include the kidneys, atypical forms of hemolytic uremic syndrome (aHUS) and membranous proliferative glomerulonephritis (MPGN), and age-related macular degeneration (AMD).
Applications
The human complement factor H ELISA kit is used for the in vitro quantitative determination of human complement factor H in serum, plasma and urine samples.
Principle
Human Complement Factor H ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 2 hours. The effective form of the plate with twelve disposable 8-well plates allows free choice of batch size for determination. Samples and standards are incubated in microtiter wells coated with antibodies that recognize human complement factor H. The peroxidase conjugate antibody will bind to the captured human complement factor H. The peroxidase conjugate 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. Draw a standard curve by plotting the absorbance (linear) relative to the corresponding concentration (logarithm) of the human complement factor H standard. The concentration of human complement factor H in a sample run at the same time as the standard can be determined from the standard curve.
Size
1 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
3.9 - 250 ng/ml
Sensitivity
3.9 ng/ml
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
100 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.

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