C5a ELISA kit [Human]-1 x 96 det. (CAT#: CB-P091-K) Datasheet

Product Type
Kit
Description
The complement system is an important factor in innate immunity. In the complement activation process of complement protein 5 (C5), C5 is proteolyzed and produces the allergic peptide C5a.
C5a is a small polypeptide composed of 74 amino acids (11kDa), derived from the N-terminus of the α chain. C5a itself has a short lifespan, and will quickly break down into more stable proteins in the serum, although it still has biological activity C5a-desArg (also known as acylation stimulating protein, ASP. Therefore, the measurement of C5a-desArg A reliable conclusion can be drawn about the level of complement activation. For convenience, the following two forms are referred to as C5a in the following.
C5a acts as an effective anaphylactoxin, causing smooth muscle contraction, vasodilation, increased vascular permeability, basophilic and mast cell degranulation, and release of lysosomal enzymes. In addition, C5a is an effective chemokine for eosinophils, basophils and monocytes.
C5a is involved in the inflammatory response seen in Gram-negative bacterial sepsis, trauma, ischemic heart disease, post-dialysis syndrome and various autoimmune diseases. Elevated C5a is associated with increased cardiovascular risk in patients with advanced atherosclerosis. Similarly, C5a is closely related to capillary leak syndrome in children with leukemia after bone marrow transplantation. C5a is also a marker in urine, which can predict the occurrence of acute transplant rejection after kidney transplantation.
The activation products of the complement cascade contain new epitopes that are not present in various natural components. The human C5a ELISA kit is based on a capture monoclonal antibody that recognizes a new epitope on C5a-desArg. This prevents cross-reaction with C5.
Applications
The human C5a ELISA kit is used for the in vitro quantitative determination of human C5a / C5a-desArg in serum and plasma.
Principle
Human C5a ELISA kit is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. The working time is 3 hours. The effective format of two plates with twelve disposable 8-well strips allows free choice of batch size for determination. The samples and standards are incubated in microtiter wells coated with antibodies that recognize human C5a. The biotinylated tracer antibody will bind to the captured human C5a. The streptavidin-peroxidase conjugate will bind the biotinylated tracer antibody. The streptavidin-peroxidase conjugate 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 C5a standard. The concentration of the human C5a sample running 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
0.3 to 20 ng/ml
Sensitivity
0.3 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 - Weak, Pig - Weak, Rabbit - Weak, Rat - Weak
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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