Quantification of cell-penetrating peptides by Fluorescence correlation spectroscopy (FCS) (CAT#: STEM-MB-1154-WXH)

Name: Quantification of cell-penetrating peptides by Fluorescence correlation spectroscopy (FCS)
SKU: STEM-MB-1154-WXH
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Introduction

Cell-penetrating peptides (CPPs) are short peptides that facilitate cellular intake and uptake of molecules ranging from nanosize particles to small chemical compounds to large fragments of DNA. The "cargo" is associated with the peptides either through chemical linkage via covalent bonds or through non-covalent interactions.
CPPs deliver the cargo into cells, commonly through endocytosis, for use in research and medicine. Current use is limited by a lack of cell specificity in CPP-mediated cargo delivery and insufficient understanding of the modes of their uptake. Other delivery mechanisms that have been developed include CellSqueeze and electroporation.

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Principle Applications Procedure Materials

Principle

Fluorescence correlation spectroscopy (FCS) is a statistical analysis, via time correlation, of stationary fluctuations of the fluorescence intensity. Its theoretical underpinning originated from L. Onsager's regression hypothesis. The analysis provides kinetic parameters of the physical processes underlying the fluctuations. One of the interesting applications of this is an analysis of the concentration fluctuations of fluorescent particles (molecules) in solution. In this application, the fluorescence emitted from a very tiny space in solution containing a small number of fluorescent particles (molecules) is observed. The fluorescence intensity is fluctuating due to Brownian motion of the particles. In other words, the number of the particles in the sub-space defined by the optical system is randomly changing around the average number. The analysis gives the average number of fluorescent particles and average diffusion time, when the particle is passing through the space. Eventually, both the concentration and size of the particle (molecule) are determined. Both parameters are important in biochemical research, biophysics, and chemistry.

Applications

• Measurement of the diffusion coefficient of biomolecules
• Detection of translational diffusions
• Measurement of the biomolecular concentration in vitro or in vivo
• Quantification of the viscosity of a solution
• Monitoring the binding or unbinding of two kinds of biomolecules
• Probing the diffusion paths of different directions and mapping the intercellular obstacles