Home \ Assays \ HNP1-3, Human, ELISA kit
Catalog # HK317

HNP1-3, Human, ELISA kit

748,001.232,00

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Human neutrophil defensins (alpha-defensins, HNP1-3) belong to the family of cationic trisulfide-containing microbicidal peptides. Besides microbicidal, the peptides exert chemotactic, immunomodulating and cytotoxic activity and participate in host defense and inflammation. Azurophilic granules of neutrophils contain Human Neutrophil Peptide (HNP)-1-4 which are highly homologous. The three principal human defensins, HNP 1-3, are unique to neutrophils and account for about 99 percent of the total defensin content of these cells. Measured amount of defensins is 3-5 mg per million human neutrophils. Activation of neutrophils leads to rapid release of defensins. Thus, only one cell type, neutrophils, may be the source of HNP1-3 measured in plasma and other body fluids during infection and inflammation. In normal plasma low levels of HNP are present ranging from undetectable level to 50-100 ng/ml, while in septic conditions the levels of HNP might be elevated to 10 mg/ml and even more. Activation of neutrophils in blood as occurs during clotting, as well as long storage of anticoagulated blood leads to a release of HNP, thus careful plasma sampling is important for possible detection of HNP. Defensins are relatively resistant to proteolysis, low pH and boiling, but have a tendency to bind to a variety of materials, including plastic and proteins. The HNP 1-3 ELISA shows cross reactivity with Rhesus monkey HNP 1-3.

Application
The human HNP1-3 (Neutrophil Defensins) ELISA kit is to be used for the in vitro quantitative determination of human HNP1-3 in plasma and cell culture supernatant samples.

Principle
The human HNP1-3 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are captured by a solid bound specific antibody. Biotinylated tracer antibody will bind to captured human HNP1-3. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate 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 HNP1-3 standards (log). The human HNP1-3 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Product type
Assays
Quantity
1 x 96 det., 2 x 96 det.
Standard range
156-10,000 pg/ml
Detection level
156 pg/ml
Working volume
100 µl/well
Species
Human
Cross reactivity
Rhesus monkey – Yes
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Precautions
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.
Disease
Autoimmunity, Infectious diseases
Assay Manual
HK317
426 kb
Safety Data Sheet

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