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AR-Red

Adenosine-Ribose Red (AR-Red) is a unique fluorescence probe that is highly specific for both aqueous Adenosine or Ribose, as compared to other sugars such as D-Glucose. D-Fructose, Galactose, Maltose, Sucrose as well as RNA and ATP. At a concentration of 4*10-9 M, AR-Red shows only a 10% fluorescence emission response to ATP as compared to the significant response (100%) to either Adenosine or Ribose, despite their similar molecular structures. AR-Red binds to either Adenosine or Ribose in a 1:1 manner.

AR-Red can be used in either an absorption or fluorescence-based manner for either the visual cellular imaging of, and/or the quantitation of either Adenosine or Ribose.  The probe can be excited over a broad wavelength range, 450 – 575 nm, with an extinction coefficient, ε = 8764 M-1 cm-1 @ 502 nm. AR-Red readily binds either Adenosine or Ribose and shows a dramatic increase in the fluorescence emission, centered at 602 nm, which is ~ 20 nm red shifted from the strong fluorescence emission spectrum of AR-Red in the absence of either Adenosine or Ribose, ~ 582 nm.

AR-Red Abs Figure 1

 

 

 

 

 

 

 

 

Figure 1. (A) Absorption spectra of AR-Red (2.3 uM) with 5.5 mM Ribose (9:1 DMSO:PBS Buffer, pH 7.4) as a function of time. (B) Absorption value at 565 nm plotted as a function of time for three diffeent concentrations of added Ribose, 0.185, 1.85 and 5.5 mM Ribose (9:1 DMSO:PBS, pH 7.4). 

 

 

 

For the quantitation of either Adenosine or Ribose, AR-Red can be used in either an Absorption or Fluorescence manner. AR-Red chelates Adenosine or Ribose in a time dependent manner, Figure 1a. Figure 1b shows the rate of change of the Absorption as monitored at 545 nm, where no further changes are observed after 30 minutes. One significant advantage of this probe, is the ability to quantitate Adenosine or Ribose. By plotting the absorption value at a fixed time point, one is readily able to quantitate the analyte, even at early times, such as 5, 10, 20 mins etc, Figure 2a. 

AR-red figure 2a

 

 

 

 

 

 

 

 

 

 

Figure 2. (A) Absorption value at 565 nm and 20 min time point plotted for 3 concentrations of Ribose, 0.185, 1.85 and 5.5 mM Ribose. (B) Fluorescence emission spectra of AR-Red both in the absence and presence of 5.5 mM Ribose, 9:1 DMSO:PBS, pH 7.4, collected after 30 min.

 

 

 

 

 

 

 

In a similar manner to Absorption, the fluorescence emission can readily be used for either visualization or quantitation, Figure 2b. The binding constant of AR-Red to Ribose is ~ 2400 M-1, measured in 9:1 DMSO/Phosphate buffer 60 Mm, pH 7.4, as compared to glucose, ~ 200 M-1, measured under otherwise identical conditions. The responses to Adenosine are almost identical to those observed for Ribose.


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