Name
Hycult Dx Complement factor H, Human, ELISA kit
Catalog nr
DH003
(lot number and expiry date are indicated on the label)
Short description
The complement system mediates a number of essential biological functions that participate in host defense against infection, initiation of the inflammatory reaction, processing and clearance of immune complexes and regulation of the immune response.
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 specifically protected against it. Each generates a C3 convertase, a serine protease that cleaves the central complement protein C3, and generates the major cleavage fragment C3b.
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 specifically protected against it. Each generates a C3 convertase, a serine protease that cleaves the central complement protein C3, and generates the major cleavage fragment C3b.
Size
1 x 96 units
Application
Manual
Description
The complement system mediates a number of essential biological functions that participate in host defense against infection, initiation of the inflammatory reaction, processing and clearance of immune complexes and regulation of the immune response.
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 specifically protected against it. Each generates a C3 convertase, a serine protease that cleaves the central complement protein C3, and generates the major cleavage fragment C3b.
Complement factor H is a relatively abundant plasma protein, with a concentration of 400-800 µg/ml, that is essential to maintain complement homeostasis and to restrict the action of complement to activating surfaces. Factor H binds to C3b, accerates the decay of the alternative pathway C3-convertase (C3bBb) and act as co-factor for the factor I-mediated proteolytic inactivation of C3b. Factor H regulates complement both in fluid phase and on cellular surfaces.
Complement protein factor H is the first regulatory protein of the alternative pathway. Factor H is a single-chain serum glycoprotein of 150 kDa with a modular structure consisting of a tandem of 20 homologous units of about 60 amino acid, called short consensus repeats (SCR).
Numerous functional sites have been identified along the 20 SCR domain structure of factor H. Three C3-binding sites have been identified; in the SCR1-4 in SCR6-10 and SCR13-20. Three polyanion binding sites like heparin and several glycoaminoglycans have also been identified in the SCR7, 13 and 20. Factor H displays anti inflammatory functions and acts as a ligand for CRP.
Factor H has two important functional domains that are located at the opposite ends of the protein. The N-terminal fragment of the factor H molecule is an essential fluid phase regulator of the alternative pathway. With the C terminal domain and SCR 7 factor H binds to cell and tissue surface and thus mediates its protective role also on host cell surface.
Genetic analyses reveal a clear association of complement factor H with different human diseases. These incude diseases of the kidney, the atypical form of Hemolytic Uremic Syndrome (aHUS) and membraneoproliferative glomerulonephritis (MPGN), and of the eye, age-related macular degeneration (AMD)
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 specifically protected against it. Each generates a C3 convertase, a serine protease that cleaves the central complement protein C3, and generates the major cleavage fragment C3b.
Complement factor H is a relatively abundant plasma protein, with a concentration of 400-800 µg/ml, that is essential to maintain complement homeostasis and to restrict the action of complement to activating surfaces. Factor H binds to C3b, accerates the decay of the alternative pathway C3-convertase (C3bBb) and act as co-factor for the factor I-mediated proteolytic inactivation of C3b. Factor H regulates complement both in fluid phase and on cellular surfaces.
Complement protein factor H is the first regulatory protein of the alternative pathway. Factor H is a single-chain serum glycoprotein of 150 kDa with a modular structure consisting of a tandem of 20 homologous units of about 60 amino acid, called short consensus repeats (SCR).
Numerous functional sites have been identified along the 20 SCR domain structure of factor H. Three C3-binding sites have been identified; in the SCR1-4 in SCR6-10 and SCR13-20. Three polyanion binding sites like heparin and several glycoaminoglycans have also been identified in the SCR7, 13 and 20. Factor H displays anti inflammatory functions and acts as a ligand for CRP.
Factor H has two important functional domains that are located at the opposite ends of the protein. The N-terminal fragment of the factor H molecule is an essential fluid phase regulator of the alternative pathway. With the C terminal domain and SCR 7 factor H binds to cell and tissue surface and thus mediates its protective role also on host cell surface.
Genetic analyses reveal a clear association of complement factor H with different human diseases. These incude diseases of the kidney, the atypical form of Hemolytic Uremic Syndrome (aHUS) and membraneoproliferative glomerulonephritis (MPGN), and of the eye, age-related macular degeneration (AMD)
Cross Reactivity
Potential cross-reacting proteins detected in the Hycult Dx Complement factor H ELISA:
Cross-reactivity for other species or proteins/peptides has not been tested.
| Cross reactant | Reactivity |
| Mouse Complement factor H | negative |
| Rat Complement factor H | negative |
| Bovine Complement factor H | negative |
| Horse Complement factor H | negative |
Cross-reactivity for other species or proteins/peptides has not been tested.
Features
- Working time of 2½ hours.
- Minimum concentration which can be measured is 3.9 ng/ml.
- Measurable concentration range of 3.9 to 250 ng/ml.
- Working volume of 100 µl/well.
Principle
- The Hycult Dx Complement factor H ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 2½ hours.
- The efficient format of 1 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 complement factor H.
- Peroxidase conjugated antibody will bind to captured human complement factor H.
- 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. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human complement factor H standards (log).
- The human complement factor H concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Use
The Hycult Dx Complement factor H ELISA kit is to be used for the in vitro quantitative determination of human complement factor H in serum, plasma and urine samples.
The results of the kit are to be used in combination with other laboratory and clinical findings for the purpose of providing information about a pathological state. The analysis should be performed by trained laboratory professionals.
The results of the kit are to be used in combination with other laboratory and clinical findings for the purpose of providing information about a pathological state. The analysis should be performed by trained laboratory professionals.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C.
Linearity
The linearity of the assay was determined by serially diluting a sample containing 250 ng/ml human factor H. The diluted samples were measured in the assay. The line obtained a slope of 0.9982 and a correlation coefficient of 1.
MSDS
| Related products | Cat # | |
|---|---|---|
| Hycult Dx Human MBL ELISA kit | DH001 | |