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Application |
for the quantitative determination of mouse Fc proteins in cell culture supernatants and serum. This assay is specific for mouse Fc, and does not cross react with human Fc. | ||||||||||
Kit components |
for 96-well plate ELISA for mouse Fc protein (2x 96-well microplates):
1 Pre-coated Microplate: 96 well polystyrene microplates coated with polyclonal antibody specific for mouse Fc protein; 2 plates 2 Detection Antibody: a horseradish peroxidase (HRP)- conjugated polyclonal antibody against mouse Fc protein; 1 vial (20 ul) 3 Mouse Fc Protein Standard: 10,000 ng/ml mouse Fc protein; 1 vial (50 ul) 4 Assay Diluent: 1x PBS with 1% BSA; 1 bottle (60 ml) 5 Wash Buffer: 10x Concentrate- PBS with detergent; 1 bottle (60 ml) 6 Color Reagent: tetramethylbenzidine (TMB); 1 bottle (20 ml) 7 Stop Solution: 0.5N hydrochloric acid solution; 1 bottle (20 ml) 8 Product Insert: product description, assay protocol, material data sheet; 1 book. |
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Background |
Fc is the tail region of an immunoglobulin G (IgG) that interacts with cell surface receptors called Fc receptors and some proteins of the complement system. The ~230 amino acid fragment generally exists as a dimer, although under reduced condition, it exists as a monomer. It is the basis of prolonged pharmacokinetics of antibodies and is commonly used as a fusion to extend half-life of fusion proteins. This assay employs the quantitative sandwich enzyme immunoassay technique. An affinity purified polyclonal antibody specific for human or mouse Fc has been coated onto a 96-well microplate. Standards, control, and samples are pipetted into the wells and any human or mouse Fc present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody specific for human or mouse Fc is added to the wells. Following an incubation and a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells. The enzyme reaction yields a blue product that turns yellow when the stop solution is added. The intensity of the color measured is in proportion to the amount of human or mouse Fc bound in the initial step. The sample values are then read off the standard curve. |
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