I am using Alexa Fluor 647 for LFIA, when i am checking the peak value for it for a specific concentration, it is variable with time. The more time it is exposed outside the less of peak i get because of which I am not able to get a proper constant result. Please help me as to what stablizer can i add while preparing various working concentrations so that it is stable for sometime
It sounds like you need and anti-fade additive. Have a look at this:
https://www.emsdiasum.com/microscopy/products/brochures/2017/EMS_Citifluor_Catalog_May2017.pdf
ATTO 647N is much more stable than Alexa Fluor 647, though ATTO 647N is very hydrophobic.
Thank you for answering Guobing Xiang
So will the hydrophobic nature hinder with its conjugation with antibody ?
Since I will be conjugating the fluorescent label (Eu3+ or Atto 647)with antibody
Shraddha
You have to try to find out, hydrophobicity will affect the water solubility, make your conjugate less water soluble!
Guobing
There are various stabilisers shown to prolong the lifetime of the fluorophore but you need to be careful about the other effects they can cause. One can describe them as antioxidants, radical scavengers or triplet state quenchers. Generally, they act as sacrificial units that are destroyed instead of the dye, so an excess is required. Probably the easiest one to use is butylated hydroxytoluene BHT.
https://pubs.rsc.org/en/content/articlelanding/2018/PP/C8PP00162F#!divAbstract
https://ui.adsabs.harvard.edu/abs/2019APS..MARA52004K/abstract
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