CircuLex Human Chitotriosidase ELISA Kit

Artikelnummer: MBL-CY-8074
Artikelname: CircuLex Human Chitotriosidase ELISA Kit
Artikelnummer: MBL-CY-8074
Hersteller Artikelnummer: CY-8074
Alternativnummer: MBL-CY-8074
Hersteller: MBL
Kategorie: Kits/Assays
Applikation: ELISA
Spezies Reaktivität: Hu
CircuLex Human Chitotriosidase ELISA Kit

The MBL Research Product CircuLex Human Chitotriosidase ELISA Kit is designed to measure the concentration of human chitotriosidase in serum.

 

Background

Chitotriosidase was first identified in the plasma of patients with Gaucher’s disease, where levels were elevated up to 1,000-fold due to lipid-laden macrophages accumulating in tissues. The enzyme, purified from spleen and cloned from human macrophages, belongs to the glycosyl hydrolase family 18 and can hydrolyze chitin. It is selectively expressed in activated tissue macrophages involved in lysosomal storage disorders, making it a useful biochemical marker of macrophage accumulation in Gaucher’s disease and for monitoring enzyme-replacement therapy. Moderate increases are also observed in other sphingolipidoses such as Niemann–Pick, GM1-gangliosidosis, and Krabbe disease.

Elevated serum chitotriosidase is additionally found in conditions associated with immune activation, including sarcoidosis, atherosclerosis, and certain infections. High enzyme activity has been detected in atherosclerotic tissue extracts and is linked to lipid-laden macrophages. It is also associated with host defense against chitin-containing pathogens and is increased in malaria. The enzyme is rapidly upregulated during immune responses, with increased mRNA and activity after stimulation by prolactin, IFN-γ, TNF, and LPS, but not IL-10.

In macrophages, chitotriosidase is synthesized as a 50-kDa protein, partly processed in lysosomes into an active 39-kDa form. A common genetic polymorphism produces a null allele that eliminates activity; about 30–40% of individuals carry this allele and approximately 6% are homozygous.

 

Principles of the Assay

The MBL Research Product CircuLex Human Chitotriosidase ELISA Kit employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for human chitotriosidase has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and the immobilized antibody binds any human chitotriosidase present. After washing away any unbound substances, an HRP conjugated antibody specific for human chitotriosidase is added to the wells. Following a wash to remove any unbound HRP conjugated antibody, the remaining conjugate is allowed to react with the substrate H2O2-tetramethylbenzidine. The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured at 450 nm. The absorbance is proportional to the concentration of human chitotriosidase. A standard curve is constructed by plotting absorbance values versus human chitotriosidase concentrations of calibrators, and concentrations of unknown samples are determined using this curve.

 

Components

  • Microplate
  • 10xWash Buffer
  • Dilution Buffer
  • Human Chitotriosidase Standard
  • HRP conjugated Detection Antibody
  • Substrate Reagent
  • Stop Solution

 

Citations

  1. Varghese AM et al. Chitotriosidase - a putative biomarker for sporadic amyotrophic lateral sclerosis. Clin Proteomics. 10, 19 (2013)(PMID:24295388)
  2. Oldoni E et al. CHIT1 at Diagnosis Reflects Long-Term Multiple Sclerosis Disease Activity. Ann Neurol. 87, 633-645 (2020)(PMID:31997416)
  3. Woollacott IOC et al. Cerebrospinal Fluid YKL-40 and Chitotriosidase Levels in Frontotemporal Dementia Vary by Clinical, Genetic and Pathological Subtype. Dement Geriatr Cogn Disord. 1-21 (2020)(PMID:32344399)
  4. Woollacott IOC et al. Cerebrospinal Fluid YKL-40 and Chitotriosidase Levels in Frontotemporal Dementia Vary by Clinical, Genetic and Pathological Subtype. Dement Geriatr Cogn Disord. 49, 56-76 (2020)(PMID:32344399)
Molekulargewicht: Dilution factors need to be taken into consideration in calculating the human chitotriosidase concentration. Results exceeding human chitotriosidase level of 3.6 ng/ml should be repeated with diluted samples.
Detektionsbereich: Dilution factors need to be taken into consideration in calculating the human chitotriosidase concentration. Results exceeding human chitotriosidase level of 3.6 ng/ml should be repeated with diluted samples.
Sensitivitaet: better than 48.3 pg/ml of sample.
Target-Kategorie: CHITO
Human Chitotriosidase Standard curve
Human Chitotriosidase concentration in serum samples