FluoBolt Noggin Fluorescence Immunoassay

Additional Information

About Metal Enhanced Fluorescence

Metal Enhanced Fluorescence (MEF) using metal nano-structures offers the possibility to increase the analytical sensitivity of systems based on Iuorescence detection dramatically. FIANOSTICS has developed a new MEF- immunoassay platform, that allows up to 300 fold gains of sensitivity. This platform is fully compatible to standard laboratory methodology using 96 well microtiter plate format and assays based on this technology can be run on any standard fIuorescence microplate reader. Its unique features enable us to develop high sensitivity, single step with no wash florescence immunoassays for low abundance biomarkers.

Assay Principle

All reagents must be at room temperature (18-26C) before use in this assay

• Add 25 μl of the selected fluorescence labeled detection antibody (DAF or DA3 or DA5 or DAA) to all wells required. Swirl gently.
• Add 20 μl of standard, control or sample to the wells according to the marked positions on the protocol sheet, swirl gently, cover tightly with the delivered adhesive film and incubate for 4 hours at 37°C, or over night at room temperature (18-26°C) in the dark.
• a)If your reader allows bottom reading, read the plate without any further processing at the Ex/Em wavelength fitting to the delivered detection antibody (495/518 nm for DAF, 550/570 nm for DA3, 650/670 nm for DA5, 679/702 nm for DAA). Gain should be set to achieve at least 10000 fluorescence units (F.U.) between the signal of the 0 pM and the 250 pM NOGGIN standard. Samples with signals exceeding the signal of the highest standard must be re-run with an appropriate dilution using sample diluent (DD).
  b)If your reader has no bottom read option or if you want to store the plate for documentation purposes, discard or aspirate the content of the wells and wash 3x with diluted wash buffer. Use a minimum of 200 µl wash buffer per well. After the final wash, remove remaining fluid by strongly tapping the plate against a paper towel. Read the plate in top configuration without any further processing at the Ex/Em wavelength fitting to the chosen detection antibody (495/518 nm for DAF, 550/570 nm for DA3, 650/670 nm for DA5, 679/702 nm for DAA). Gain should be set to achieve at least 10000 fluorescence units (F.U.) between the signals of the 0 pM and the 250 pM NOGGIN standard. Samples with signals exceeding the signal of the highest standard must be re-run with appropriate dilution using sample diluent (DD).
  Quality of bottom reading (3a) may vary between microplate readers. For first time users we suggest to perform the washing step and follow protocol 3b (ED).
• Store the plate with desiccant at 4°C (2-8°C) in the aluminium bag. Unused wells are stable until expiry date stated on the label. Fluorescence signals of standards, controls and samples remain detectable for at least two month at the plate surface, depending on signal intensity achieved.