Ideal for situations where a rapid, simple and sensitive assay solution is desired for quantitative analyses
These assays employ highly validated antibodies that have been screened to isolate the enzyme in active form and in complexes that likely represent their associations in situ.
Advantages of dipstick assays
This technique requires minimal sample preparation and few overall steps. The total assay time is less than one hour and requires as little as 3µg of sample material. It is suitable for a wide range of sample types: such as tissue, cultured cells, blood and cheek swab material and can be used in human, mouse and rats species and does not require isolation of mitochondria. There are corresponding quantitative sandwich ELISA assays for most enzyme activity assays and combining the two, it is possible to generate specific activity data.
Dipstick enzyme activity assays apply a novel approach, whereby target proteins are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurs. Assay are also available in a multiplex format for multiple target proteins.