Published TCIMAIL newest issue No.197
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Europium Fluorophore DTBTA-Eu3+-labeled Secondary Antibodies and Streptavidin
-Highly-sensitive Detection Probes by Time-resolved Fluorometry-
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Fig: SDS-Page of S0993 and G0505(reducing conditions) |
Exitation and emission spectra of DTBTA-Eu3+ in 0.05M borate buffer at pH9.0 (1.5x10-6M) |
Advantages
No cross talk of excitation light
Stable fluorescence in various aqueous buffers
Long fluorescent life time (τ = 1.02 ms)
• 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
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.