Human A1AT Lateral Flow Kit
For Research Use Only
B.I.T.S. Human A1AT Lateral Flow Kit
Urine & Serum
0.78 – 50ng/mL
Intra-assay CV% < 15.3
Inter-assay CV % < 15
Average 89% Range 76-97%
Average 94% Range 81-117%
1 in 20, 1 in 40, 1 in 80, with recovery < 117%
1 in 200K, 1 in 400K, 1 in 800K, with recovery < 103.2%
Human Alpha-1 antitrypsin (A1AT)
The unique B.I.T.S.® Lateral Flow Kit for Human Alpha-1 antitrypsin (A1AT) is the first lateral flow assay product to be developed by Mologic. It provides fast, accurate assays of A1AT in plasma and urine.
The B.I.T.S.® A1AT Lateral Flow Assay needs no sample work-up and delivers a result in just 10 minutes.
Refined and simplified step-wise procedure minimises operator error and significantly reduces time between sampling and results. Time saved in executing the assay will give you more time to do more research.
Kits are manufactured to the highest quality by Mologic scientists who have developed these assays from scratch. This ensures stability and batch to batch reproducibility better than industry standards. Despite this commitment to quality, the price of the kits is in line with the competition – quality without compromise.
The B.I.T.S.® A1AT Lateral Flow Kit is the first reliable, rigorously validated assay for human urinary and serum A1AT of its kind, either reported in the literature or available commercially. This is an important product milestone, which will open up the use of A1AT as a robust biomarker with great utility in inflammatory disease research.
B.I.T.S.® Immunoassays – a great new resource for Biomedical and life sciences researchers working in inflammation and host immune response to infection!
B.I.T.S.® – our lateral flow assay range will be expanding throughout 2015.
A1AT Lateral Flow Assay Format.
Alpha-1 antitrypsin (A1AT) is a 52kDa glycoprotein belonging to the serpin family of protease inhibitors.
A1AT is produced by the liver and secreted into the bloodstream.
The balance between A1AT and proteases contributes to the integrity of the extracellular matrix and is important in the regulation of tissue integrity.
When present in the sample, A1AT binds to anti-A1AT mouse fab complexed to colloidal gold, which is then captured by a second anti-A1AT mouse fab which has been immobilised in a line across the cassette membrane, producing a visible red line. A procedural control is included to indicate that the assay has been performed correctly.
Example of a an A1AT standard curve.
Equilibrate all materials and prepared reagents to room temperature (18-25°C) prior to use. It is recommended to assay all standards, controls and samples in duplicate.
1. Equilibrate all materials and prepared reagents to room temperature (18-25˚C) prior to use.
2. Prepare samples as directed in the previous sections.
3. Remove the lateral flow device from its pouch and place it on a flat surface and test within 10 minutes of opening..
4. Add 80μL of diluted sample to the round well at the base of the device using a pipette.
5. Read the test line result against the score card exactly 10 minutes after adding the sample to the device.
6. Any trace of a pink line in the test area indicates a positive result. Any results read outside 10 minutes should be considered invalid and must be repeated.
Summary of the assay procedure.
A1AT Score Card.
A1AT is a major protease inhibitor that controls tissue degradation. Circulating levels of A1AT in the blood of healthy individuals are typically between 1.0 and 2.7 mg/mL [Donato 2012] but are dramatically elevated in acute and chronic inflammation in order to reduce host tissue damage caused by inflammatory proteases. A reduction of A1AT levels can cause a change in collagen metabolism. A1AT deficiency is an uncommon genetic disease that can lead to emphysema, hepatitis, cirrhosis, and chronic obstructive pulmonary disease (COPD) [de Serres 2014].
A1AT covalently binds to proteases and causes irreversible inhibition.
In oxidative stress, methionine residues of A1AT can become oxidised, which leads to polymerization and subsequent inactivation.
Mologic’s B.I.T.S.® A1AT LF kit is only able to detect free, monomeric A1AT.
NEGLIGIBLE CROSS-REACTIVITY TO VARIOUS COMPOUNDS
The specificity of the assay was evaluated by measuring the degree of cross-reactivity of various compounds that were used in place of A1AT. The amount of antibody that was bound at each concentration was calculated and the approximate percentage of cross reactivity was calculated from the amount of compound that produced a signal equivalent to 50%.
L J Donato, S M Jenkins, C Smith, J A Katzmann & M R Snyder (2012).
Reference and Interpretive Ranges for α1-Antitrypsin Quantification by Phenotype in Adult and Pediatric Populations.
American Journal of Clinical Pathology, 138, 398-405.
F de Serres & I Blanco (2014)
Role of alpha-1 antitrypsin in health and disease.
Journal of Internal Medicine. 276(4), 311-335.