Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IgG has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any IgG present is bound by the immobilized antibody and the detection antibody specific for IgG is binds to the combination of capture antibody-IgG in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IgG present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IgG standard dilutions and IgG sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures .
Immunoglobulin G (IgG) is a type of antibody. Representing approximately 75% of serum antibodies in humans, IgG is the most common type of antibody found in blood circulation. IgG molecules are created and released by plasma B cells. Each IgG has two antigen binding sites. IgG are involved in the regulation of allergic reactions. According to Finkelman, there are two pathways of systemic anaphylaxis: antigens can cause systemic anaphylaxis in mice through classic pathway by cross-linking IgE bound to the mast cell receptor FcεRI, stimulating the release of both histamine and platelet activating factor (PAF). In the alternative pathway antigens form complexes with IgG, which then cross-link macrophage receptor FcγRIII and stimulates only PAF release. Human IgG is the major immunoglobulin in blood and plays a key role in the humoral immune response.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with antibody to human IgG-HRP (812)||1 plate|
|Human IgG standard lyophilized, 50000 pg/ml upon reconstitution||2 vials|
|Antibody to human IgG-HRP solution||2 vials|
|Standard /sample Diluent||4 bottles|
|Antibody to human IgG-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 390pg/mL.|
|ASSAY RANGE||780 - 50000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human IgG.|