ANCA-C (PR3) ELISA Kit

Name: ANCA-C (PR3) ELISA Kit
SKU: DEIA3141
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The ANCA-C (PR3) ELISA Kit is a test system for the quantitative measurement of IgG class autoantibodies against proteinase 3 (PR3) in human serum or plasma.

Contents of Kit

1. Divisible microplate: Consisting of 12 modules of 8 wells each. Ready to use. 96 (8×12) wells
2. Calibrators A-F (0, 5, 10, 20, 40, 100 U/mL), containing serum/buffer matrix (PBS, BSA, detergent, NaN₃ 0.09%), yellow. Ready to use. 1.5 mL × 6
3. Control positive (1) and negative (2), containing PR3 antibodies in a serum/buffer matrix (PBS, BSA, detergent, NaN₃ 0.09%), yellow, Ready to use. The concentration is specified on the certificate of analysis. 1.5 mL × 2
4. Sample buffer, containing PBS, BSA, detergent, preservative NaN3 0.09%, yellow, 5× conc. 20 mL
5. Enzyme conjugate; containing anti-human IgG antibodies, HRP labelled; PBS, BSA, detergent, preservative ProClin 300 0.05%, light red. Ready to use. 15 mL
6. TMB substrate; containing 3,3',5,5'-Tetramethylbenzidin, colorless. Ready to use. 15 mL
7. Stop solution; contains acid. Ready to use. 15 mL
8. Wash Buffer, containing Tris, detergent, preservative NaN₃ 0.09%; 50× conc. 20 mL

Storage

Store the kit at 2-8°C in the dark. Do not expose test reagents to heat, sun or strong light during storage and usage. Store microplate sealed and desiccated in the clip bag provided. Unopened reagents are stable until expiration of the kit. See labels for individual batch. Diluted Wash solution and Sample Buffer are stable for at least 30 days when stored at 2-8°C. We recommend consumption on the same day.

Performance Characteristics

Calibration
This assay system is calibrated in relative arbitrary units, since no international reference preparation is available for this assay.
Expected values
In a normal range study with samples from healthy blood donors the following ranges have been established with this ELISA assay: Cut-off 5 U/mL

Detection Range

The calculation range of this ELISA assay is 0-100 U/mL

Detection Limit

Functional sensitivity was determined to be: 0.5 U/mL

General Description

Anti-neutrophil cytoplasmic antibodies (ANCA) are diagnostic markers for ANCA-associated vasculitides. Anti-PR3 characterizes granulomatosis with polyangiitis (GPA, formerly: Wegener's granulomatosis). The test supports the differential diagnosis of vasculitis when used in combination with other laboratory and clinical findings.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Novel SNPs in the CD18 gene validate the association with MPO-ANCA(+) vasculitis

GENES AND IMMUNITY

Authors: Meller, S; Jagiello, P; Borgmann, S; Fricke, H; Epplen, JT; Gencik, M

Abstract

Wegener granulomatosis (WG), microscopic polyanglitis (MP), and Churg-Strauss syndrome (CSS) are characterized by the presence of anti-neutrophil cytoplasmic antibodies (ANCA). Anti-myeloperoxidase (MPO)-ANCA are a typical feature of MP and CSS, while anti-proteinase 3 (PRTN3)-ANCA are highly specific for WG. Several reports indicate that ANCA may directly contribute to pathological processes, ie, through an increase of adhesivity between polymorphonuclear (PMN) and endothelial cells (EC). PMN interact and endothelium interact via the adhesion cascade (AC). CD18 is a key molecule of the AC, as CD18 defects abbrogate the adhesion of PMN and cause leukocyte adhesion deficiency, an immunodeficient trait. We have screened the entire coding and regulatory regions of the CD18 gene. Ten single nucleotide polymorphisms (SNP) were identified, four of them showing significant associations with MPO-ANCA(+) vasculitis. One of these SNPs was localized in an alternate transcription initiation site. This polymorphism may influence CD18 gene expression, resulting in dose-dependent increase in adhesion and consecutively facilited degranulation and respiratory burst. In this manner the pro-adherent genotype may predispose to MPO-ANCA(+) vasculitis.

Sequencing and bioinformatics analysis of the differentially expressed genes in herniated discs with or without calcification

INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE

Authors: Shao, Jia; Yu, Miao; Jiang, Liang; Wu, Fengliang; Liu, Xiaoguang

Abstract

The purpose of this study was to detect the differentially expressed genes between ossified herniated discs and herniated discs without ossification. In addition, we sought to identify a few candidate genes and pathways by using bioinformatics analysis. We analyzed 6 samples each of ossified herniated discs (experimental group) and herniated discs without ossification (control group). Purified mRNA and cDNA extracted from the samples were subjected to sequencing. The NOISeq method was used to statistically identify the differentially expressed genes (DEGs) between the 2 groups. An in-depth analysis using bioinformatics tools based on the DEGs was performed using Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and protein-protein interaction network analysis. The top 6 DEGs were verified using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). A total of 132 DEGs was detected. A total of 129 genes in the ossified group were upregulated and 3 genes were found to be downregulated as compared to the control group. The top 3 cellular components in GO ontologies analysis were extracellular matrix components. GO functions were mainly related to the glycoprotein in the cell membrane and extracellular matrix. The GO process was related to completing response to stimulus, immune reflex and defense. The top 5 KEGG enrichment pathways were associated with infection and inflammation. Three of the top 20 DEGs [sclerostin (SOST), WNT inhibitory factor 1 (WIF1) and secreted frizzled related protein 4 (SFRP4)] were related to the inhibition of the Wnt pathway. The ossified discs exhibited a higher expression of the top 6 DEGs [SOST, joining chain of multimeric IgA and IgM (IGJ; also known as JCHAIN), defensin alpha 4 (DEFA4), SFRP4, proteinase 3 (PRTN3) and cathepsin G (CTSG)], with the associated P-values of 0.045, 0.000, 0.008, 0.010, 0.015 and 0.002, respectively, as calculated by the independent sample t-test. The gene expression profiling of the 2 groups revealed differential gene expression. Thus, our data suggest that Wnt pathway abnormality and local inflammation may be related to disc ossification.