Abstract:
:The single-molecule fluorescence technique is becoming a general and mature tool to probe interactions and dynamics of biomolecules with ultra high precision and accuracy. However, nonspecific adsorption of biomolecules to the flow cells remains a major experimental riddle for the study of many complex biological systems, especially those exhibiting low binding affinity and presenting with weakly populated intermediates. Many novel surface passivation methods have been introduced to reduce nonspecific interactions. Here, we present an effective and inexpensive method to significantly reduce nonspecific binding of biomolecules in conventional poly (ethylene glycol) (PEG)-based surface passivation protocols, without additional exogenous effects. In particular, we propose a simple 10 min Tween-20 treatment for the PEG passivated surface, which could further increase the hydrophilicity of the surface and thus promote passivation efficacy by about 5 to 10 times. We anticipate that this new procedure will find broad practical applications and extend the current reaches of single-molecule fluorescence studies.
journal_name
Phys Bioljournal_title
Physical biologyauthors
Pan H,Xia Y,Qin M,Cao Y,Wang Wdoi
10.1088/1478-3975/12/4/045006subject
Has Abstractpub_date
2015-06-29 00:00:00pages
045006issue
4eissn
1478-3967issn
1478-3975journal_volume
12pub_type
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