Description
The ImmuChrom Alpha-1 Antitrypsin ELISA Kit is intended for the quantitative determination of alpha-1-antitrypsin (A1AT) in human stool or serum samples.
Alpha-1-Antitrypsin is a 52 kD glycoprotein, which is produced by the liver, intestinal macrophages, monocytes and mucous membrane cells of the gut. It belongs to the group of acute phase proteins and is one of the most important proteinase inhibitor. Alpha-1-antitrypsin inhibits, beside others, the proteinases trypsin and the elastase of neutrophiles. A lack of α-1-AT leads to an enhanced proteolysis. Only a very small amount of alpha-1-antitrypsin is cleaved or resorbed in the gut. Therefore the measurement of α-1-AT in stool reflects the permeability of the gut during inflammatory processes.
Type
Sandwich ELISA, HRP-labelled antibody
Other names
A1AT ELISA, α-1-AT, SERPINA1, SERPIN A1, serum trypsin inhibitor, alpha 1 proteinase inhibitor, Alpha-1-antiproteinase, fecal Alpha-1-Antitrypsin, faecal Alpha-1-Antitrypsin
Principle of Method
The α-1-AT ELISA test determines human alpha-1-antitrypsin according to the “sandwich”-principle. α-1-AT in sample, standard and controls binds to antibodies, which are coated to the microtiterplate. After a washing step a peroxidase labeled detection antibody is added. A second washing step is followed by the addition of the substrate which is converted to a colored product by the peroxidase. The reaction is terminated by the addition of an acidic stop solution. The optical densities are read at 450 nm (against the reference wavelength 620 nm) in a microtiterplate reader. The α-1-AT concentration can be calculated from the standard curve.
Sample Types
Stool, Serum
Sample Volume
- Stool: 100 mg (collected sample)
- Serum: 0.0004 µL (actual amount of sample loaded in well after dilution)
Standard Curve
Calibration Range: 3.3 ng/ml - 90 ng/ml
The example curve given above is only for demonstration.
Limit of Detection
0.3 ng/ml
For the determination of the detection limit 20 replicates of the standard 0 were measured. After addition of the 3 fold standard deviation to the mean value the concentration was read from the standard curve.
Precision & Reproducibility
Intended Use
This assay is intended for research use only and is not for use in diagnostic procedures.
Resources
Product References
- ARUP Laboratories validation poster: Performance Evaluation of a Polyclonal Based Fecal Alpha-1-Antitrypsin ELISA
- ARUP Laboratories validation study: Performance Evaluation of an ELISA for the Quantitative Measurement of α1-Antitrypsin in Stool
- Shivakumar, Nirupama et al. “Tryptophan oxidation in young children with environmental enteric dysfunction classified by the lactulose rhamnose ratio.” The American journal of clinical nutrition vol. 116,4 (2022): 970-979. doi:10.1093/ajcn/nqac171
- Bhuiyan, Taufiqur R et al. “Mucosal-Associated Invariant T (MAIT) cells are highly activated in duodenal tissue of humans with Vibrio cholerae O1 infection: A preliminary report.” PLoS neglected tropical diseases vol. 16,5 e0010411. 12 May. 2022, doi:10.1371/journal.pntd.0010411
- Sinha, Bireshwar et al. “Effect of Community-Initiated Kangaroo Mother Care on Fecal Biomarkers of Gut Function in Low Birth Weight Infants in North India: A Randomized Clinical Trial.” The American journal of tropical medicine and hygiene vol. 106,3 945-952. 20 Dec. 2021, doi:10.4269/ajtmh.21-0877
- Zambrana, Luis E et al. “Rice bran supplementation modulates growth, microbiota and metabolome in weaning infants: a clinical trial in Nicaragua and Mali.” Scientific reports vol. 9,1 13919. 26 Sep. 2019, doi:10.1038/s41598-019-50344-4
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Havt, Alexandre et al. “Prevalence and virulence gene profiling of enteroaggregative Escherichia coli in malnourished and nourished Brazilian children.” Diagnostic microbiology and infectious disease vol. 89,2 (2017): 98-105. doi:10.1016/j.diagmicrobio.2017.06.024
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Ilex Life Sciences LLC is an official distributor of Immuchrom GmbH.