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Europium Fluorophore DTBTA-Eu3+-labeled Secondary Antibodies and Streptavidin
-Highly-sensitive Detection Probes by Time-resolved Fluorometry-

     

 

Fig: SDS-Page of S0993 and G0505(reducing conditions)
S0993: Detect major band that corresponded to Streptavidin DTBTA-Eu3+.
G0505: Detect 2 major bands that corresponded to the heavy chains and light chains of Goat Anti Mouse IgG DTBTA-Eu3+.

 

Exitation and emission spectra of DTBTA-Eu3+ in 0.05M borate buffer at pH9.0 (1.5x10-6M)
The excitation spectrum is right and the emission spectrum is left.


 

Advantages

No cross talk of excitation light
・λex, max = 335 nm
・λem, max = 616 nm
Sharpened emission spectrum
Large Stokes shift (the difference in wavelength between positions of the band maxima of the absorption and emission spectra)

Stable fluorescence in various aqueous buffers
Available in Tris, TE, PBS, etc. for wide use

Long fluorescent life time (τ = 1.02 ms)
Time-resolved fluorometric measurement can remove background fluorescence from the sample matrix and often gives detectability better than one order of magnitude compared to those of conventional fluorometric assays.

Comparison of secondary antibody conjugated to DTBTA-Eu3+ or FITC

<Assay condition>
Dilute the Mouse IgG to each concentration. Coat 96-well plates with diluted Mouse IgG. Block the plates with BSA/TBST. Incubate with Goat Anti-Mouse IgG Conjugates prepared from DTBTA or FITC at 2.5 μg/mL. After incubation, measure the fluorescence intensity on a plate reader.
DTBTA-Eu3+; excitation=340 nm, emission=620 nm. Lag Time : 450 μsec
FITC; excitation=485 nm, emission=520 nm.