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Complement C5 ELISA kit [Human]-2 x 96 det. (CAT#: CB-P072-K) Datasheet

Product Type
Kit
Description
Complement C5 (molecular weight is about 190kDa) is a key component of the complement system. This system plays an important role in both innate and adaptive immune responses, and can produce inflammatory and protective responses to pathogen attacks before adaptive responses occur. The complement system consists of a complex family of proteins and receptors, which are found in the circulatory system, tissues and other body fluids. Excessive/uncontrolled activation/regulation of the complement system involves many diseases, such as asthma, lupus erythematosus, glomerulonephritis, various forms of arthritis, autoimmune heart disease, multiple sclerosis, inflammatory bowel disease, Paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome and ischemia-reperfusion injury and transplant rejection. Three main pathways within the complement system have been described, namely the classical pathway, the alternative pathway and the lectin pathway, which are activated by different stimuli. The classical pathway is triggered by immune complexes. Through the lectin pathway of surface-bound lectins; and protect AP through all unprotected surfaces. All three pathways converge on complement C3. The cleavage of C3 induces the cleavage of C5, which in turn leads to the production of membrane attack complex (MAC). This protein complex is the ultimate common effector responsible for lysing susceptible microorganisms or destroying cells. Mutations in the C5 gene can cause a deficiency of complement component 5. This disease is characterized by repeated bacterial infections.
Applications
The Human C5 ELISA kit is used for the in vitro quantitative determination of C5 in plasma and serum samples.
Principle
The Human C5 ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle, with a working time of 3.5 hours.
The effective form of the plate with twelve disposable 8-well plates allows free choice of batch size for determination.
The samples and standards are incubated in microtiter wells coated with antibodies that recognize human C5.
The biotinylated tracer antibody will bind to the captured human C5.
 Streptavidin peroxidase conjugate will bind to the biotinylated tracer antibody.  Streptavidin-peroxidase conjugate will react with the substrate tetramethylbenzidine (TMB).  The enzyme reaction can be 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 C5 standard.
The concentration of human C5 in the sample that was performed at the same time as the standard can be determined from the standard curve.
Size
2 x 96 det.
Detection Method
Enzyme immunoassay (ELISA) technique
Detection Limit
3.1 to 200 ng/ml
Sensitivity
3.1 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
Cross Reactivity
Horse - No, Mouse - No, Pig - No, Rat - No
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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