Description
Intended Uses: This Nucleo-h IgG ELISA is a solid phase enzyme immunoassay with human native nucleosomes isolated from the eukaryotic cell line HeLa for the quantitative and qualitative detection of antibodies against nucleosomes and their components dsDNA and histones in human serum.
Principle of the Assay: Serum samples diluted 1:101 are incubated in the microplates coated with the specific antigen. Antibodies, if present in the specimen, bind to the antigen. The unbound fraction is washed off in the following step. Afterwards anti-human immunoglobulins conjugated to horseradish peroxidase (conjugate) are incubated and react with the antigen-antibody complex of the samples in the microplates. Unbound conjugate is washed off in the following step. Addition of TMB-substrate generates an enzymatic colorimetric (blue) reaction, which is stopped by diluted acid (color changes to yellow). The intensity of color formation from the chromogen is a function of the amount of conjugate bound to the antigen-antibody complex and this is proportional to the initial concentration of the respective antibodies in the sample.
Background: Nucleosomes are the basic structure of chromatin important for the compaction of DNA in the nucleus. A nucleosome comprises of four histones H2A, H2B, H3 and H4 forming an octamer and 146 bp of dsDNA wrapped 1 ¾ turns around the core (H2A, H2B, H3, H4)2 octamer. Histone H1 interacts with the nucleosome and together with linked-DNA connects neighboring nucleosomes