5-Carboxytetramethylrhodamine succinimidyl ester

Use of sulfo-NHS-LC-SPDP or other heterobifunctional crosslinkers to modify PAMAM dendrimers may be done along with the use of a secondary conjugation reaction to couple a detectable label or another protein to the dendrimer surface. Patri et al. (2004) used the SPDP activation method along with amine-reactive fluorescent labels (FITC or 6-carboxytetramethylrhodamine succinimidyl ester) to create an antibody conjugate, which also was detectable by fluorescent imaging. Thomas et al. (2004) used a similar procedure and the same crosslinker to thiolate dendrimers for conjugation with sulfo-SMCC-activated antibodies. In this case, the dendrimers were labeled with FITC at a level of 5 fluorescent molecules per G-5 PAMAM molecule. 

This approach has been shown to work with a number of different fluorescent probes such as the short-wavelength fluorophores dansyl sul-fonyl chloride and coumarin chloride and the long-wavelength fluorophores tetramethylrhodamine-5-(and-6)-isothiocyanate [5(6)-TRITC], 5-(and-6)-carboxytetramethylrhodamine, succinimidyl ester [5(6)-TAMRA, succin-imidyl ester] and lissamine rhodamine B sulfonyl chloride (each in conjunction with different binding functionalities on the SAM surface. 

Fig. 4.9 Synthesis scheme for the preparation of the fluorescent SAMs (DA, CA, TMA, DS, CS, TMS, DU, CU, and TMU) on glass and silicon surfaces, (i) Red Al, toluene, 45°C (ii) dansyl chloride, 7-dimethylaminocoumarin-4-acetic acid succinimidyl ester, or 5-(and-6)-carboxytetramethylrhodamine, succinimidyl ester, acetonitrile, rt (iii) benzenesulfonyl chloride orp-isopropylphenyl isocyanate, acetonitrile, rt. 36 Reproduced with permission of The Royal Society of Chemistry...

Chang and Lin [50] determined vigabatrin in human plasma by capillary electrophoresis and laser-induced fluorescence after precolumn derivatization with 5-carboxytetramethylrhodamine succinimidyl ester. Optimal separation and detection were obtained with an electrophone buffer of 50 mM sodium borate (pH 9.5) containing 10 mM sodium dodecyl sulfate and a green He-Ne laser with fluorescence detection at 589 nm and excitation at 543 nm. The assay was rectilinear over the concentration range of 1.5-200 /iM and the lower limit of detection was 0.13 /iM. Both the... 

Using this approach, a CE-SDS method with LIF detection has been developed and validated as a quality control procedure for the purity determination of a recombinant mAb [125]. 5-Carboxytetramethylrhodamine succinimidyl ester was used as fluorescent reagent and the optimal conditions for derivatization of the nonreduced and reduced (reacted with dithiothreitol) mAb and... 

Fluorescein tags (Fig. 3) Carboxytetramethylrhodamine succinimidyl ester from Molecular Probes (Eugene, OR and Leiden, The Netherlands) and sulforhodamine from Sigma-Aldrich, St. Louis, MO. 

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