人抗組蛋白IgG檢測試劑盒說明書

Human Anti histones IgG ELISA kit

Cat. No. 3300-130-HNG

Intended Use:

Anti-Histone is an indirect solid phase enzyme immunoassay (ELISA) for the measurementof IgG class autoantibodies to histone in human serum or plasma. The assay is intendedfor research use only.

Introduction:

Histones are highly alkaline proteins found in eukaryotic cell nuclei that package and orderthe DNA into structural units called nucleosomes. They are the chief protein componentsof chromatin, acting as spools around which DNA winds, and play a role in gene regulation.Without histones, the unwound DNA in chromosomes would be very long (a length to widthratio of more than 10 million to 1 in human DNA). Histones are found in the nuclei ofeukaryotic cells, and in certain Archaea, namely Euryarchaea, but not in bacteria. Theunicellular algae known as dinoflagellates are the only eukaryotes that are known tocompletely lack histones. Five major families of histones exist: H1/H5, H2A, H2B, H3, andH4. Their molecular weights range from 11 to 21 kDa. They contain a lot of basic aminoacid residues which presumably interact with the negative charged groups of DNA.Furthermore they contain polar amino acid residues which may be important for theirinteraction among each other. Histones H2A, H2B, H3 and H4 are known as the corehistones, while histones H1 and H5 are known as the linker histones. Two of each of thecore histones assemble to form one octameric nucleosome core particle, and 147 basepairs of DNA wrap around this core particle 1.65 times in a left-handed super-helical turn.The linker histone H1 binds the nucleosome at the entry and exit sites of the DNA, thuslocking the DNA into place[7] and allowing the formation of higher order structure.All of the nucleic acids (DNA) of eukaryotic cells are associated with proteins. The complexstructure of DNA and its associated small basic proteins, which are called histones, isknown as chromatin. Both, the histones comprise about 50 percent of the total mass ofeukaryotic chromosomes. The complex of DNA and the histones can be dissociated bytreatment of the eukaryotic chromatin with salt or diluted acids. Five different types ofhistones are known. They are called H1, H2A, H2B, H3 and H4. In correlation to theirfundamental function in the organization of chromatin, the structure of all histones in alleukaryotes is highly conserved.. Antibodies to histones usually produce a homogeneous,rim or speckled pattern of nuclear staining in indirect immunofluorescenc.

Sensitivity

The lower detection limit for Anti-histones IgG has been determined at 1.0 U/ml.SpecificityThe microplate is coated with histones highly purified by affinity chromatography.The Anti- histones test kit is specific only for autoantibodies directed to histones.No cross reactivities to the other ENA or DNA antigens have been observed.

Calibration

Since no international reference preparation for anti-histone autoantibodies isavailable, the assay system is calibrated against the WHO reference preparationfor human anti nuclear factor (homogenous), MRC 66/233. With the Anti-Histonekit this preparation is determined at a concentration of 100 U/ml.

LIMITATIONS OF PROCEDURE

The Anti-histones IgG ELISA is a diagnostic aid. A definite clinical diagnosisshould not be based on the results of a single test, but should be made by thephysician after all clinical and laboratory findings have been evaluated.

INTERFERING SUBSTANCES

No interference has been observed with haemolytic (up to 1000 mg/dL), lipemic(up to 3 g/dL triglycerides) or bilirubin (up to 40 mg/dL) containing sera. Nor haveany interfering effects been observed with the use of anticoagulants. However forpractical reasons it is recommended that grossly hemolysed or lipemic samplesshould be avoided.

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