The Biocolor Fastin Elastin Assay is a quantitative, colourimetric method designed for the precise measurement of elastin in biological samples. This kit, manufactured by Biocolor Ltd and supplied by companies such as Ilex Life Sciences LLC and AkrivisBio, is a tool primarily used in research and clinical diagnostics rather than a consumer product. It is employed to analyse elastin content in tissues, fluids, and cell culture media, providing critical data for studies in developmental biology, pathology, and tissue engineering. The assay operates on a dye-binding principle, where elastin forms a complex with a specific dye (TPPS) that can be measured spectrophotometrically. This overview details the kit's methodology, suitable applications, and technical specifications, drawing exclusively from the provided source material. It is intended for researchers and laboratory personnel in the UK seeking to understand the assay's capabilities and limitations.
The Fastin Elastin Assay is a quantitative, dye-binding method for analysing elastins released into tissue culture medium and extracted from biological materials. The dye label employed is 5,10,15,20-tetraphenyl-21H,23H-porphine tetra-sulfonate (TPPS). This method is designed to measure various elastin forms, including soluble tropoelastins, lathyrogenic elastins, and insoluble elastins after solubilisation to elastin polypeptides such as α-elastin and κ-elastin. The dye reagent binds to the basic and non-polar amino acid sequences found in mammalian elastins. The principle of the method involves several key steps. First, incubation of samples containing soluble elastin with the Fastin Dye Reagent causes an elastin-dye complex to form, which is an insoluble complex that then precipitates. Second, the dye-labelled elastin is isolated by centrifugation, and the unbound dye is removed. The elastin-bound dye is then eluted and measured spectrophotometrically. Third, the elastin content of unknown samples is calculated by comparison against a calibration curve prepared using a standard comprising water-soluble elastin, which is supplied with the kit. The signal is directly proportional to the elastin content, and the assay is noted for being simple, reliable, and selective with a linear range of 0 μg-40 μg elastin per well.
The assay is suitable for a range of sample types. For in-vivo applications, it can be used on tissues and fluids. However, insoluble elastin requires conversion to water-soluble α-elastin using the oxalic acid reagents and extraction protocol supplied with the kit. For in-vitro applications, the assay can directly measure soluble elastin secreted into cell culture medium. It is important that samples for analysis are free from any particulate material, such as cell debris or insoluble extracellular matrix material. The presence of other soluble proteins or complex carbohydrates does not interfere with the Fastin Elastin Assay. Tropoelastin is the elastin form typically found in cell-culture medium during in vitro cell culture. However, a specific note indicates that elastin in conditioned cell media may be below the detection limit of the kit. The kit can also quantify various elastin forms, spanning from immature tropoelastin to mature, insoluble elastin fibres.
The technical specifications of the Biocolor Fastin Elastin Assay, as detailed in the provided sources, include the following. The detection method is colourimetric, with measurements taken at 513 nm. The assay range is 0 to 500 µg/ml, with a limit of detection of 50 µg/ml. The assay run-time is 4 hours. The kit provides for 110 measurements in total, which allows a maximum of 48 samples to be run in duplicate alongside a standard curve. For the unopened kit, storage is recommended at room temperature. The sensitivity of the assay is reported as 5 µg. The manufacturer is Biocolor Ltd.
The relevance of elastin measurement is underscored by its role in normal development and organ function. Impairment of elastin synthesis or the proteolytic degradation of elastin fibres can result in major clinical pathologies. For instance, mutations in the Elastin gene are linked to diseases such as Williams-Beuren syndrome, Cutis laxa, and Supravalvular aortic stenosis. Furthermore, several skin diseases have been associated with abnormalities of elastin, and marked fragmentation of elastin is observed in pulmonary emphysema. The Fastin Assay, therefore, serves as a valuable tool for research into these conditions, enabling the quantification of elastin to better understand disease mechanisms and potential therapeutic interventions.
In summary, the Biocolor Fastin Elastin Assay is a well-defined, quantitative colourimetric assay for measuring elastin in biological samples. Its methodology is based on a specific dye-binding principle, and it is designed for use with both soluble and insoluble elastin (after appropriate sample preparation). The kit provides a standard curve for quantification and is suitable for a variety of sample types, including tissues, fluids, and cell culture media. Its specifications, such as the assay range, detection limit, and measurement capacity, are clearly outlined. The assay's importance is highlighted by the clinical significance of elastin in various pathologies. For researchers in the UK, this kit offers a straightforward solution for elastin quantification in a laboratory setting.