Anti-Nucleosomes Antibody ELISA Kit

Name: Anti-Nucleosomes Antibody ELISA Kit
SKU: DEIA-JY2407
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The kit is intended for quantitative determination of IgG class autoantibodies against Nucleosomes in human serum.

Contents of Kit

Reagents that need to be prepared:
1. 1 bottle of 5× Sample buffer, 20 mL - 5 times concentrated (white cap: yellow solution)
Contains: tris (Tris), sodium chloride (NaCl), bovine serum albumin (BSA), azide Sodium <0.1% (preservative)
2. 1 bottle of 50× Washing buffer, 20 mL - 50 times concentrated (white cap: green solution)
Contains: Tris, sodium chloride (NaCl), Tween 20, stack Nitrogen sodium﹤0.1% (preservative)

Ready-to-use reagents:
1. Negative quality control 1 bottle, 1.5 mL (green cap: colorless solution)
Contains: diluted human serum, sodium azide <0.1% (preservative)
2. Positive quality control 1 bottle, 1.5 mL (red cap: yellow solution)
Contains: diluted human serum, sodium azide <0.1% (preservative)
3. Cut-off quality control 1 bottle, 1.5 mL (blue cap: yellow solution)
Contains: diluted human serum, sodium azide <0.1% (preservative)
4. Calibrator 6 bottles, each bottle is 1.5 mL, respectively 0, 3, 10, 30, 100, 300 U/mL
(Color deepens with concentration: yellow solution)
Contains: diluted human serum, sodium azide <0.1% (preservative)
5. Enzyme conjugate 1 bottle, 15 mL IgG (blue cap: blue solution)
Contains: Horseradish peroxidase-labeled anti-human immunoglobulin
6. Tetramethylbenzidine (TMB) substrate 1 bottle, 15 mL (black cap)
Contains: Stabilized Tetramethylbenzidine (TMB)/Hydrogen Peroxide (H₂O₂)
7. Stop solution 1 bottle, 15 mL (white cap: colorless solution)
Contains: 1 mol/L (M) hydrochloric acid
8. Divisible microplate consisting of 12 modules of 8 wells each, coated with native human Nucleosomes. Ready to use; 1 plate

Storage

1. Store the kit at 2°C - 8 °C.
2. Keep microplate wells sealed in a dry bag with desiccants.
3. The reagents are stable until expiration of the kit.
4. Do not expose test reagents to heat, sun or strong light during storage and usage.
5. Diluted sample buffer and wash buffer are stable for at least 30 days when stored at 2°C - 8 °C

Performance Characteristics

1. Analytical sensitivity
After testing the sample buffer of this kit 30 times, the analytical sensitivity of this kit is 1.0 U/mL.
2. Specificity and sensitivity
The microplate is coated with highly purified natural human nucleosomes, and no cross-reactivity with other autoantigens has been found.
The diagnostic specificity is 90% and the sensitivity is 84-88%.
3. Response linearity
The good linearity of this kit test was demonstrated by serial dilutions of selected sera. However, due to the presence of naturally occurring heterogeneous autoantibodies, some samples may not comply with this rule.

Precision

Intra-assay Precision (Precision within an assay) C.V < 10%.
Inter-assay Precision (Precision between assays) C.V < 10%.

General Description

Nucleosomes are the basic nuclear chromosomal structures that keep DNA in the nucleus compact. The nucleosome is composed of four histamine complexes: H2A, H2B, H3 and H4, and forms a 146bp 8-mer double-stranded DNA. H1 serves as a link between nucleosomes and other nucleosomes linked to DNA.
Anti-double-stranded DNA (dsDNA) and anti-histamine complex antibodies are common features of systemic lupus erythematosus (SLE). In 1993, Burlingame et al. first discovered anti-chromatin antibodies corresponding to T cell-mediated autoantibody immune responses in a mouse model. This study shows that the first autoimmune response in a mouse model of systemic wolf erythematosus (SLE) is directly anti-nucleosome. Therefore, anti-nucleosome-specific antibodies are specific markers for early systemic lupus erythematosus (SLE), which can recognize the antigenic determinants of natural nucleosome molecules. The subsequent autoimmune response is divided into antinucleosome, anti-DNA, and anti-histamine complexes.
The detection rate of nucleosome-specific antibodies in patients with systemic lupus erythematosus (SLE) is 84 to 88%.

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